(2022) 20:291
Osoro et al. BMC Medicine
https://doi.org/10.1186/s12916-022-02498-8
RESEARCH ARTICLE
Open Access
Prevalence of microcephaly and Zika virus
infection in a pregnancy cohort in Kenya, 2017–
2019
Eric Osoro1,2* , Irene Inwani3, Cyrus Mugo3, Elizabeth Hunsperger4, Jennifer R. Verani4, Victor Omballa5,
Dalton Wamalwa3, Chulwoo Rhee6, Ruth Nduati3, John Kinuthia7, Hafsa Jin8, Lydia Okutoyi9,
Dufton Mwaengo10, Brian Maugo3, Nancy A. Otieno5, Harriet Mirieri1, Mufida Shabibi11, Peninah Munyua4,
M. Kariuki Njenga1,2 and Marc‑Alain Widdowson4,12
Abstract
Background: Zika virus (ZIKV), first discovered in Uganda in 1947, re‑emerged globally in 2013 and was later associ‑
ated with microcephaly and other birth defects. We determined the incidence of ZIKV infection and its association
with adverse pregnancy and fetal outcomes in a pregnancy cohort in Kenya.
Methods: From October 2017 to July 2019, we recruited and followed up women aged ≥ 15 years and ≤ 28 weeks
pregnant in three hospitals in coastal Mombasa. Monthly follow‑up included risk factor questions and a blood sample
collected for ZIKV serology. We collected anthropometric measures (including head circumference), cord blood,
venous blood from newborns, and any evidence of birth defects. Microcephaly was defined as a head circumference
(HC) < 2 standard deviations (SD) for sex and gestational age. Severe microcephaly was defined as HC < 3 SD for sex
and age. We tested sera for anti‑ZIKV IgM antibodies using capture enzyme‑linked immunosorbent assay (ELISA) and
confirmed positives using the plaque reduction neutralization test (PRNT90) for ZIKV and for dengue (DENV) on the
samples that were ZIKV neutralizing antibody positive. We collected blood and urine from participants reporting fever
or rash for ZIKV testing.
Results: Of 2889 pregnant women screened for eligibility, 2312 (80%) were enrolled. Of 1916 recorded deliveries,
1816 (94.6%) were live births and 100 (5.2%) were either stillbirths or spontaneous abortions (< 22 weeks of gesta‑
tion). Among 1236 newborns with complete anthropometric measures, 11 (0.9%) had microcephaly and 3 (0.2%)
had severe microcephaly. A total of 166 (7.2%) participants were positive for anti‑ZIKV IgM, 136 of whom became
seropositive during follow‑up. Among the 166 anti‑ZIKV IgM positive, 3 and 18 participants were further seroposi‑
tive for ZIKV and DENV neutralizing antibodies, respectively. Of these 3 and 18 pregnant women, one and 13 (72.2%)
seroconverted with antibodies to ZIKV and DENV, respectively. All 308 samples (serum and urine samples collected
during sick visits and samples that were anti‑ZIKV IgM positive) tested by RT‑PCR were negative for ZIKV. No adverse
pregnancy or neonatal outcomes were reported among the three participants with confirmed ZIKV exposure. Among
newborns from pregnant women with DENV exposure, four (22.2%) were small for gestational age and one (5.6%) had
microcephaly.
*Correspondence: eric.osoro@wsu.edu
1
Washington State University Global Health Kenya, One Padmore Place,
George Padmore Road, Off Ngong Road, Nairobi, Kenya
Full list of author information is available at the end of the article
© The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which
permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the
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Osoro et al. BMC Medicine
(2022) 20:291
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Conclusions: The prevalence of severe microcephaly among newborns in coastal Kenya was high relative to pub‑
lished estimates from facility‑based studies in Europe and Latin America, but little evidence of ZIKV transmission.
There is a need for improved surveillance for microcephaly and other congenital malformations in Kenya.
Keywords: Zika virus, Microcephaly, Pregnancy, Kenya
Background
In 2015, the Asian lineage of Zika virus (ZIKV), a mosquito-borne pathogen first described in Uganda in 1947,
spread globally to cause widespread outbreaks in the
Americas and Asia. Although more than 80% of these
ZIKV infections were mild or subclinical, infections in
pregnant women were associated with multiple congenital fetal malformations collectively referred to as congenital Zika syndrome and include microcephaly, congenital
contractures, brainstem dysfunction, and severe cerebral
and eye lesions [1–4].
In February 2016, the World Health Organization
declared the ZIKV outbreak a public health event of
international concern, and calls were made for pregnancy cohort studies to better understand the role of
clinical and subclinical infections in congenital malformations and characterize the incidence and spectrum of
these adverse outcomes [5]. Subsequently, prospective
studies in Latin America during the outbreak reported a
wide range of seropositivity to ZIKV infection (8–53%)
and confirmed the association between Zika infection in
pregnant women and adverse fetal outcomes [1, 6, 7].
In sub-Saharan Africa (sSA), two outbreaks of ZIKV
(Asian lineage) associated with microcephaly were
reported in Cape Verde and Angola in 2016 [8, 9].
Although modeling studies suggest that the risk of transmission of flaviviruses, including ZIKV, dengue, and yellow fever, is high in many areas of sSA [10], limited data
exist on ZIKV circulation in this region. The surveillance
of microcephaly in the continent is very rudimentary,
and it remains unclear if ZIKV infection has caused an
undocumented burden of microcephaly and other birth
defects [11]. One recent analysis of specimens from a
dengue outbreak has shown that ZIKV co-circulated in
Kenya in 2013, while another study found a prevalence of
neutralizing anti-ZIKV antibodies of up to 7% in Northern Kenya [12, 13]. However, cross-reacting antibodies
between flaviviruses can complicate the interpretation of
these findings [14]. Moreover, serologic and viral surveillance cannot easily differentiate between the Asian and
African lineages of Zika. Recent studies suggest that the
African ZIKV lineage virus has higher transmissibility
and pathogenicity compared to the Asian lineage strain,
and infection in pregnant women may be more likely to
cause total fetal loss than congenital deformities associated with the Asian lineage [15].
We established a prospective cohort study among
pregnant women in an area of coastal Kenya predicted
to be at risk for circulating ZIKV [16] because of circulating dengue and the Aedes aegypti vector, the primary
mosquito that transmits ZIKV. We aimed to determine
the incidence and seroprevalence of ZIKV infection and
assess its association with adverse pregnancy outcomes.
Methods
Study setting
We enrolled pregnant women who sought care at one of
three hospitals in Mombasa on the east coast of Kenya.
These were (a) Coast General Hospital, the second largest
public hospital in Kenya with a capacity of 700 beds; (b)
Port Reitz Hospital with a capacity of 166 beds; and (c)
Bomu Hospital, a private health hospital with a capacity
of 45 beds, and an HIV care reference center.
Participants and eligibility criteria
Pregnant women presenting for antenatal care (ANC)
were eligible for enrolment if they met the following
criteria: (1) aged 15 years or older, (2) had a confirmed
pregnancy from ANC records or ultrasound, (3) had an
estimated gestational age ≤ 28 weeks, and (4) planned
to attend ANC and deliver at the study hospitals. We
excluded women who were found to have an ectopic or
molar pregnancy by ultrasound findings and were participating in trials of experimental drugs and devices or
those incarcerated.
Study procedures
After informed consent and enrolment, a baseline questionnaire was administered, a dating ultrasound was conducted for those without an obstetric ultrasound before
enrolment, and 5 ml of venous blood sample was collected. When ultrasound was not available, the date of
the last menstrual period, antenatal medical records on
gestational age, or clinical assessment of fundal height
were used in that order to assess gestational age. Participants were followed up monthly at the study clinic for
the duration of their pregnancy at which time a questionnaire on risk factors was completed and a 5-ml blood
sample collected. Participants were also asked to report
and attend the clinic for assessment if they experienced
any fever or rash. If symptom onset was within 7 days of
the report, urine and blood samples were collected.
Osoro et al. BMC Medicine
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At delivery, maternal, placenta, and cord blood samples were collected, and a questionnaire administered.
The study staff conducted a newborn physical assessment within 48 h of birth for gross abnormalities and
measured birth weight, head circumference, and other
anthropometric characteristics. Additionally, 3 ml of
venous blood was drawn from the newborn (Fig. 1).
Participants who did not deliver at the study sites were
asked to attend the study facilities within 2 weeks for
maternal and newborn blood sample collection.
Data collection
During enrolment and monthly follow-up, structured
questionnaires were administered by trained study
staff to collect data on sociodemographic characteristics, pregnancy characteristics, potential risk factors
for congenital defects, and environmental risk factors
for ZIKV infection such as water storage, exposure to
mosquitoes, and prevention against mosquito bites.
Other data collected included medical and obstetric
history, signs and symptoms of possible ZIKV infection, and other relevant infections, including details
and timing in relation to pregnancy/gestational age
and pregnancy outcomes (such as live birth, miscarriage, and stillbirth). Study data were collected via
electronic tablets using REDCap [17]. We abstracted
data from participants’ ANC, sick visits, and delivery
medical records to capture relevant information. Clinical measurements of the newborn were performed following standard procedures by study staff at the study
sites [18].
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Laboratory procedures
All samples were barcoded, stored at − 20 °C, and
shipped to the Kenya Medical Research Institute/Centers
for Disease Control and Prevention laboratory in Nairobi
for storage at − 80 °C until tested in the same laboratory.
Maternal and newborn sera were tested for anti-ZIKV
IgM antibodies using the ZIKV IgM antibody capture
enzyme-linked immunoassay (MAC-ELISA) as described
previously [19]. Briefly, Immunol plates were coated with
anti-human IgM (Kirkegaard and Perry Laboratories)
and incubated overnight before incubation for 2 h with
patient sera diluted at 1:400. Vero-cell-derived ZIKV E6
antigen was added to the plate and detected using 6B6C1
anti-ZIKV IgG horseradish peroxidase-conjugated monoclonal. Due to the potential of cross-reactivity with
other flaviviruses (particularly dengue virus), anti-ZIKV
IgM-positive samples were confirmed using the ZIKV
90% plaque reduction neutralization test (PRNT90) as
previously described [20, 21]. Furthermore, samples
with ZIKV PRNT90 titers ≥ 1:20 were tested for dengue
virus-2 (DENV2, used because it cross-reacts with other
DENV serotypes) by PRNT90 (Fig. 2). Urine and serum
samples from participants with possible ZIKV infection
(defined below) and anti-ZIKV IgM positive were tested
for ZIKV RNA using CDC real-time reverse transcription polymerase chain reaction (rRT-PCR).
Study definitions
A possible case of ZIKV infection was defined as any
participant with fever (≥ 38° C) or a history of fever
(within the previous 7 days) or a rash. A probable case
was defined as the presence of IgM antibodies against
ZIKV. A confirmed case of ZIKV infection was defined
Fig. 1 Schedule of study procedures in the pregnancy cohort, Kenya, 2017–2019
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Fig. 2 Testing algorithm for Zika virus antibodies among participants in the pregnancy cohort, Kenya, 2017–2019. DENV, dengue fever virus; ZIKV,
Zika virus; PRNT, plaque reduction neutralization assay
as detection of ZIKV RNA by rRT-PCR in serum or
urine samples, or ZIKV IgM positive and PRNT90
for ZIKV with titer ≥ 20 and ZIKV PRNT90 titer
ratio ≥ fourfold higher when compared to DENV [22].
A confirmed DENV-positive case was defined as DENV
PRNT90 titer ratio ≥ fourfold higher when compared to
ZIKV PRNT90 titers (Fig. 2).
Microcephaly was defined as a head circumference < 2 standard deviations (SD) below the mean
for sex and gestational age and severe microcephaly
as a head circumference < 3 SD based upon INTERGROWTH-21st standards [18]. We defined stillbirth
as a baby born without any sign of life and birthweight of ≥ 500 g or, if missing, ≥ 22 completed weeks
of gestation. Abortion was defined as pregnancy
losses < 22 weeks of gestation. Low birth weight was
defined as < 2500 g. Small-for-gestational-age (SGA)
was defined as a birth weight z-scores of < − 1.28 at
birth (equivalent to the 10th percentile) and extreme
SGA as a birth weight of < − 1.88 z-scores (equivalent
to the 3rd percentile) [18].
Sample size
The sample size estimation was based on the hypothesis that pregnant women with incident ZIKV infection
(primary exposure) have a higher risk of microcephaly
and other congenital malformations (primary outcome)
in their offspring at delivery. Assuming a background
risk of microcephaly of 0.03%, a minimum detectable relative risk of 25, and the proportion of the study
cohort with incident Zika virus infection of 3 to 5%
[23], a sample size of between 2127 and 2669 pregnant
women was needed to achieve a power of at least 80%.
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Data analysis
Results
Descriptive analyses were performed using categorical
and continuous variables, compared using the chi-square
test or Fisher’s exact test or using the Student t-test.
Continuous variables with a non-normal distribution
were compared using the Kruskal–Wallis test. For all the
analyses, data were considered significant at a p-value
of < 0.05. All data were analyzed using the R statistical
software [24].
Characteristics at enrolment and follow‑up
Ethical considerations
This study was approved by the Kenyatta National Hospital/University of Nairobi Ethical Review Committee
[P71/102/2017], the Washington State University institutional review board [IRB No. 15897], and the CDC institutional review board [# 7021]. Participants (those below
18 years were considered emancipated minors) provided
informed consent at the time of recruitment.
Fig. 3 Flow diagram of the pregnancy cohort, Kenya, 2017–2019
From October 2017 to July 2019, a total of 3069 pregnant women were referred from the ANC clinics and
2889 (94.1%) were screened for eligibility. Of these,
2647 (91.6%) were eligible and 2312 (87.3%) enrolled in
the study. Among enrollees, delivery outcome data were
available for 1916 (82.9%) while 396 (17.1%) were either
lost to follow-up (delivery outcome not recorded) or
withdrew from the study (Fig. 3). Participants who were
lost to follow-up or withdrew from the study were significantly older and with higher gestational age at enrolment
(Table S1).
The mean age of the enrolled participants was
28.4 years (SD 5.6), median gestational age of 20.0 weeks
[interquartile range (IQR) 15.1–24.1] at enrollment. A
total of 2028 (87.5%) participants had a dating ultrasound
completed. Overall, 343 (15.9%) of the participants were
HIV infected, and of these, 267 (77.8%) were enrolled
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at Bomu hospital, and 310 (92%) were on antiretroviral medication. Nearly 30% of the participants reported
a previous diagnosis of chikungunya or dengue by a
health care worker. Other sociodemographic and obstetric characteristics are included in Table 1. The median
number of follow-up visits among participants was four
(IQR = 2–5). One hundred and four women reported
fever (n = 99) or rash (n = 10) during pregnancy with 20
identified at enrolment and 84 during follow-up. Of all
the symptomatic pregnant women, blood and urine samples were obtained from 81 (77.9%).
Table 2 Delivery and newborn characteristics of the pregnancy
cohort, Kenya, 2017–2019
Characteristic
Total n (%)
Delivery outcome
1916
Live newborn
1816 (94.8)
Stillbirth
66 (3.4)
Abortion (< 22 weeks)
34 (1.8)
Birth weight categories
1432
< 2500 g
83 (5.8)
2500–4000 g
1272 (88.8)
> 4000 g
77 (5.4)
Newborn anthropometric measures
Delivery outcomes
Of 1916 recorded delivery outcomes, 1816 (94.8%)
were live births (including 25 (1.3%) twin gestations)
and 100 (5.2%) deliveries were either stillbirths or abortions (Table 2). More than two-thirds [1365 (71.2%)] of
the deliveries were at the study facilities. A total of 23
Microcephaly (head circumference < 2SD)
1236
Severe microcephaly (head circumference < 3SD)
1236
3 (0.2)
Small for gestational age (birth weight < 1.8SD)
1200
142 (11.8)
1200
57 (4.8)
Extremely small for gestational age (birth
weight < 2.8SD)
11 (0.9)
SD standard deviation
Table 1 Sociodemographic, obstetric, and clinical characteristics
of the pregnancy cohort at enrolment, Kenya, 2017–2019
Characteristic
N
Age, years mean (SD)
2312
Education level completed
2278
28.4 (± 5.6)
Primary and below, n (%)
592 (26.0)
Secondary, n (%)
1016 (44.6)
Tertiary, n (%)
Occupation
670 (29.4)
2312
Employed, n (%)
953 (41.2)
Self‑employed, n (%)
366 (15.8)
Unemployed, n (%)
992 (42.9)
Gestational age in weeks, median [IQR]
2312
Previous pregnancies
2309
0, n (%)
20.0 [15.1–24.1]
691 (29.9)
1 to 3, n (%)
1415 (61.3)
Above 3, n (%)
203 (8.8)
Previous pregnancies’ outcome
Laboratory results
1618
Pregnancy loss
528 (32.6)
Newborn with birth defects
19 (1.2)
Any chronic diseasea, n (%)
2312
143 (6.2)
Systolic blood pressure (mmHg)b, mean (SD)
2309
109.3 (11.7)
Diastolic blood pressure (mmHg)c, mean (SD)
2307
67.9 (9.3)
Hemoglobin level (g/dl)d, mean (SD)
2145
11.2 (2.7)
HIV infected, n (%)
2160
343 (15.9)
HIV infected and using ARVs, n (%)
343
310 (92.3)
Positive VDRL for syphilis, n (%)
2163
29 (1.3)
IQR interquartile range, SD standard deviation, VDRL Venereal Disease Research
Laboratory, ARV antiretroviral drugs
a
Chronic disease included asthma, hypertension, diabetes, and epilepsy
b
Normal range 90–120
c
Normal range 60–80
d
Normal range 9.5–15
(1.2%) of all newborns had at least one overt congenital
abnormality: 12 (52.2%) congenital umbilical hernias,
6 (20.7%) upper and 4 (13.8%) lower limb deformities,
and one Down syndrome. One newborn had both upper
and lower limb deformities. Among the 66 stillbirths, 19
(28.7%) were examined at delivery with no gross anomalies noted.
Among 1236 (64.5%) newborns with data on sex and
gestational age and head circumference measurements,
11 (0.9%) presented with microcephaly and 3 (0.2%) with
severe microcephaly. This translates to a prevalence of 90
(95% CI 45–159) cases of microcephaly per 10,000 births
and 20 (95% CI 5–71) cases per 10,000 births for severe
microcephaly. Slightly over 10% of the newborns were
small for gestational age (SGA) (Supporting information).
We collected and tested a median of 4 (IQR 2–5) serum
samples per participant, for a total of 8276 samples
including 6518 (78.8%) maternal, 924 (11.2%) newborn,
and 834 (10.1%) cord blood samples. During the study, 81
women presented with fever or rash. All serum and urine
samples collected within 7 days of onset were negative by
RT-PCR for ZIKV and by MAC-ELISA for IgM.
Samples from 2293 (99.2%) were tested for anti-ZIKV
IgM, and 166 (7.2%) women had at least one blood
sample positive for anti-ZIKV IgM. Among them, 131
(78.9%) tested positive in one sample, 25 (15.1%) in two
samples, and 10 (6%) in more than two samples for a total
of 213 positive IgM samples. The majority (136 [81.9%])
of these participants were anti-ZIKV IgM negative at
enrolment and became seropositive during the follow-up
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period corresponding to an anti-ZIKV IgM incidence of
5.9 per 100 pregnancies. Two seropositive participants
reported a history of clinical diagnosis of dengue fever
within 4 weeks before collection of the sample that was
anti-ZIKV IgM positive.
The proportion of anti-ZIKV IgM-seropositive participants who had newborns with microcephaly compared
to seronegative participants was not statistically different
(1.6% vs 0.8%, p-value = 0.682, n = 1236). Similarly, the
proportion of SGA among anti-ZIKV IgM seropositive
compared to seronegative participants was not statistically different (15.4% vs 11.4%, p-value = 0.245, n = 1200)
(Table 3). There was no statistical association between
ZIKV IgM seroconversion and microcephaly or SGA.
Among the 166 participants with anti-ZIKV IgM,
samples from 144 (86.7%) participants were tested by
PRNT and 3 (2.1%) were confirmed ZIKV positive and
18 (12.5%) confirmed DENV positive while 7 (4.9%)
were inconclusive, for a seroprevalence of 0.1% for
ZIKV in the cohort (n = 2293). None of the newborns
from the three ZIKV PRNT-positive women had microcephaly. Of the three participants positive for ZIKV by
PRNT, two were positive at enrolment. The pregnancy
and delivery characteristics of the three participants
are outlined in Table 4.
Among the 18 participants who were DENV PRNT
positive, four tested positive at two different study
visits. Thirteen (72.2%) of these 18 participants were
DENV confirmed during the follow-up period while
five (27.8%) participants were confirmed DENV positive at enrolment. Four (22.2%) of the DENV-positive
participants delivered small-for-gestational-age offspring and one (5.6%) participant delivered a newborn
with microcephaly (the participant was DENV PRNT
positive at enrolment at 11.2 weeks of gestation). Seven
(31.8%) of the participants with confirmed DENV were
HIV infected and one participant had a stillbirth at
31.3 weeks of gestation.
We found the sera of one mother and her newborn
collected during delivery to be seropositive for antiZIKV IgM and DENV PRNT. Another mother had
maternal serum collected at delivery that was antiZIKV IgM positive but ZIKV PRNT negative (therefore
Table 3 Comparison of pregnancy and newborn characteristics by anti‑Zika virus IgM positivity in the pregnancy cohort, Kenya,
2017–2019
Characteristic
Total
Anti‑ZIKV IgM
p‑value
Positive
Negative
Age in years at enrolment, median [IQR]
2293
26.7 [23.6–31.6]
28.1 [24.2–32.4]
0.158
Gestational age in weeks at enrolment, median [IQR]
2293
19.5 (14.0–23.0)
20.0 (15.1–23.9)
0.153
Small‑for‑gestational‑age newborns, n (%)
1200
19 (15.4)
123 (11.4)
0.245
Microcephaly in newborns, n (%)
1236
2 (1.6)
9 (0.8)
0.682
IQR interquartile range, ZIKV Zika virus
Table 4 Pregnancy and delivery characteristics of three participants of a pregnancy cohort with confirmed ZIKV by PRNT, Kenya,
2017–2019
Characteristic
Participant 1
Participant 2
Participant 3
No. of samples positive/tested by ELISA for anti‑
ZIKV IgM
3/5
2/6
3/6
No. of samples positive/tested by PRNT
1/3
2/2
1/3
IgM positive at enrolment
Yes
Yes
No
Gestational age at PRNT confirmation
• Enrolment at 12.3 weeks
• Enrolment at 13.9 weeks
• Follow‑up at 24.8 weeks
• Follow‑up at 33.3 weeks
Gestational age at enrolment
12.3 weeks
13.9 weeks
14.3 weeks
Age in years at enrolment
30.1
36.9
21.3
HIV infection
Negative
Positive
Negative
Pregnancy outcome
Lost to follow‑up
Live newborn
Live newborn
Gestational age at delivery
Lost to follow‑up
37.3
41.1
Small‑for‑gestational‑age newborn
Lost to follow‑up
No
No
DENV dengue fever virus, ZIKV Zika virus, PRNT plaque reduction neutralization assay
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not evaluated for DENV) and newborn serum that was
anti-ZIKV IgM and DENV PRNT positive.
Discussion
In this large longitudinal study of pregnant women in
coastal Kenya from 2017 to 2019, we found little evidence of active Zika virus circulation, but possible evidence of a higher prevalence of severe microcephaly of
0.2% compared with 0.13% expected from a reference
population based on INTERGROWTH-21st standards [18]. However, the prevalence of any microcephaly
in our study was less than half of the expected (2.3%) in
the reference population. We found evidence of ZIKV
neutralizing antibodies in only three among 2293 participants screened first for IgM. Two of the participants
were already seropositive for ZIKV by IgM and PRNT
at enrolment between 12 and 14 weeks of gestation, and
therefore, it remains unclear if the infection occurred
during pregnancy or previously. However, of note, 136
of 166 women became IgM positive during follow-up of
which 18 were determined positive for DENV by PRNT.
With the assumption that the IgM and ZIKV PRNT positivity reflected cross-reaction of dengue IgM antibodies,
these results suggest substantial dengue circulation in
this population, but since only the ZIKV PRNT positive
were tested for DENV, it remains likely that the true burden of dengue was higher. The higher DENV seropositivity reflects high transmission of DENV in an area that has
reported regular outbreaks over the last decade [25, 26].
Almost three-quarters of the participants with evidence
of exposure to DENV seroconverted during the study follow-up, further evidence of the high level of transmission
of DENV.
The prevalence of microcephaly of 90 cases per 10,000
births in our study was about half that reported in a
retrospective study (200 cases per 10,000 births) from
an earlier period (2012–2016) in Kilifi, coastal Kenya
[27]. However, the level of microcephaly reported in
our study was at least 9 times higher than < 10 cases
per 10,000 births reported from studies in Europe and
Latin America [28, 29]. It was also 1.5 times higher than
reported in Brazil (60 cases per 10,000 births) at the peak
of the ZIKV outbreak in 2016. However, a study in China
reported a higher prevalence of 410 cases per 10,000
births [30]. None of the newborns with microcephaly in
our study had evidence of ZIKV and only one had confirmed DENV exposure. Nonetheless, the low number of
infections in our study means that we cannot exclude an
association. Microcephaly is associated with infectious
(such as rubella, cytomegalovirus, herpes simplex virus,
immunodeficiency virus, toxoplasmosis, and syphilis),
environmental, and genetic causes. The higher prevalence of microcephaly in coastal Kenya as reported in our
Page 8 of 11
study and from the study in Kilifi [27] compared to facility-based studies from other countries could be because
the cited studies used clinical data, in addition, to head
circumference measurements to define microcephaly and
variations in the head circumference cutoff for defining
microcephaly (some studies defined microcephaly head
circumference < 3 SD below the mean for sex and gestational age).
The low levels of ZIKV seropositivity in our study are
similar to findings from several studies in Africa and
Asia. A study in a long-term cohort on HIV transmission among women in Mombasa assessed by PRNT for
anti-ZIKV neutralizing antibodies among febrile presentations found ZIKV seropositivity in one of 900 participants [31]. Similar findings with fewer than five
participants seropositive for ZIKV from among several
hundred tested were reported in Uganda, DR Congo,
Cambodia, and Vietnam [21, 32–34]. Commonly in
these studies, DENV seropositivity was higher than that
of ZIKV. Contrary to our findings, a study in northern
Kenya reported a high ZIKV seroprevalence of 7% and
DENV seroprevalence of 1%, although it was not clear
how the potential cross-reactivity was assessed [12].
The low levels of ZIKV transmission could be due to
the competition of the virus on the same vector and host.
In countries that reported high ZIKV seroprevalence of
50–73% during the 2013–2016 outbreak, the incidence
of DENV decreased during the ZIKV outbreak but
increased after the outbreak [35]. Additionally, the differences in transmission could be a reflection of the cyclic
variation of flavivirus transmission or other favorable climatic conditions [36]. Recently, it has been shown that
different subspecies of Aedes aegypti may have different
susceptibility to ZIKV infection and that the vectors in
the Americas represent a population of mosquitoes susceptible to ZIKV infection, whereas similar mosquitoes
in Africa may be resistant [37].
Our findings of IgM positivity in neonatal sera suggest
prepartum infection and vertical transmission of DENV
since IgM antibodies do not cross the placenta. Vertical
transmission of DENV is associated with preterm deliveries and low birth weight although the mechanism is not
clear. Congenital malformations of the nervous system
have also been associated with DENV infection in pregnancy [38–40].
Our study had several limitations. First, the screening antibody test was ELISA for ZIKA IgM and we did
not undertake DENV IgM testing. We may have underestimated the true ZIKV seroprevalence because we
only included ZIKV-positive sera from those that were
positive by IgM and PRNT and without high titers of
DENV antibodies. Some of the sera initially positive for
ZIKV by PRNT and then judged as dengue infections
Osoro et al. BMC Medicine
(2022) 20:291
Page 9 of 11
because of high DENV titer may have been acute ZIKV
infections but with high DENV background neutralizing antibodies due to historic DENV infections. Second, since we were not able to detect any ZIKV virus,
we were unable to determine if any circulating ZIKV
was from the African lineage or the Asian lineage
that spread throughout the Americas and was associated with microcephaly. Third, we did not test systematically for anti-ZIKV IgG which would have indicated
true seroprevalence in the cohort and the pre-existing
immunity to ZIKV infection. However, the low yield
from the ZIKV PRNT overall testing suggests that the
background ZIKV seroprevalence is low. Fourth, our
assessment of adverse neonatal outcomes was only
conducted during delivery. Some of the malformations
associated with congenital Zika syndrome become
apparent during the months after delivery which our
study could not establish because of a lack of infant follow-up. Finally, about one-third of the deliveries were
not within the study facilities and we, therefore, could
not assess some of the potential adverse outcomes
among the newborns. In addition, almost two-thirds of
the stillbirths were not examined for congenital abnormalities because the deliveries were not within the
study facility, or the bodies were disposed of before the
study staff could examine them.
Acknowledgements
We thank the Ministry of Health, the County Government of Mombasa, Kenya
Medical Research Institute, the hospital administration of Coast General, Port
Reitz and Bomu hospitals, study staff, and the participants for their role and
support in the implementation of the study.
Conclusions
The epidemiology and transmission of ZIKV in Africa
remain unclear. However, we found no evidence of substantial ZIKV transmission in coastal Kenya, and most
of the population of coastal Kenya, including pregnant
women, are susceptible to infection. Our study highlights the need for enhanced surveillance of arboviral
infections and congenital abnormalities in Kenya and
throughout Africa. Understanding transmission, lineages of ZIKV, and vector presence and susceptibility is
key to risk assessment and future prevention and control of outbreaks.
Consent for publication
Not applicable.
Abbreviations
ANC: Antenatal care; CDC: US Centres for Disease Control and Prevention;
DENV: Dengue virus; ELISA: Enzyme‑linked immunosorbent assay; HC: Head
circumference; IgM: Immunoglobulin M; IQR: Interquartile range; PRNT: Plaque
reduction neutralization assay; rRT‑PCR: Real‑time reverse transcription poly‑
merase chain reaction; SD: Standard deviation; SGA: Small‑for‑gestational‑age;
sSA: Sub‑Saharan Africa; ZIKV: Zika virus.
Supplementary Information
The online version contains supplementary material available at https://doi.
org/10.1186/s12916‑022‑02498‑8.
Additional file 1: Table S1. Comparison of some characteristics between
participants who completed follow‑up and thoselost to follow‑up.
Disclaimer
The findings andconclusions in this manuscript are those of the authors and
do not necessarilyrepresent the official position of the US Centers for Disease
Control andPrevention or the Government of Kenya.
Authors’ contributions
Conceptualization: EO, II, CM, EH, JV, DW, RN, JK, LO, DM, NO, KN, MAW. Formal
analysis: EO, II, CM, KN, MAW. Funding acquisition: KN, MAW. Methodology: EO,
II, CM, EH, JV, VO, DW, CR, RN, JK, HJ, LO, DM, BM, NO, MS, PM, KN, MAW. Project
administration: EO, II, CM, HM, KN, MAW. Supervision: EO, II, EH, RN, HS, HM, MS,
KN, MAW. Writing — original draft: EO, II, CM, KN, MAW. The authors read and
approved the final manuscript.
Funding
This work was funded with funds from the Centers for Disease Control and
Prevention, Cooperative Agreement No. U01GH002143‑01.
Availability of data and materials
The datasets used and/or analyzed during the current study are available from
the corresponding author on reasonable request.
Declarations
Ethics approval and consent to participate
This study was reviewed and approved by the Kenyatta National Hospital/Uni‑
versity of Nairobi Ethical Review Committee [P71/102/2017], the Washington
State University institutional review board [IRB No. 15897], and the US CDC
institutional review board [# 7021]. All participants (those below 18 years
were considered emancipated minors) provided informed consent before
enrolment.
Competing interests
The authors declare that they have no competing interests.
Author details
1
Washington State University Global Health Kenya, One Padmore Place,
George Padmore Road, Off Ngong Road, Nairobi, Kenya. 2 Paul G. Allen School
of Global Health, Washington State University, Pullman, USA. 3 Department
of Pediatrics and Child Health/Kenyatta National Hospital, University of Nai‑
robi, Nairobi, Kenya. 4 Division of Global Health Protection, Centers for Disease
Control and Prevention, CDC Kenya, Nairobi, Kenya. 5 Center for Global Health
Research, Kenya Medical Research Institute, Nairobi, Kenya. 6 Division of Global
Health Protection, CentersforDiseaseControlandPrevention, Atlanta, USA.
7
Research and Programs Department, Kenyatta National Hospital/University
of Nairobi, Nairobi, Kenya. 8 Coast General Hospital, Mombasa, Kenya. 9 Depart‑
ment of Obstetrics and Gynecology/Kenyatta National Hospital, University
of Nairobi, Nairobi, Kenya. 10 Institute of Tropical and Infectious Diseases,
University of Nairobi, Nairobi, Kenya. 11 Port Reitz Hospital, Mombasa, Kenya.
12
Institute of Tropical Medicine, Antwerp, Belgium.
Received: 10 February 2022 Accepted: 25 July 2022
References
1. Sanchez Clemente N, Brickley EB, Paixão ES, De Almeida MF, Gazeta RE,
Vedovello D, et al. Zika virus infection in pregnancy and adverse fetal
outcomes in São Paulo State, Brazil: a prospective cohort study. Sci Rep.
2020;10:12673. https://doi.org/10.1038/s41598‑020‑69235‑0.
Osoro et al. BMC Medicine
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
(2022) 20:291
Pomar L, Malinger G, Benoist G, Carles G, Ville Y, Rousset D, et al. Asso‑
ciation between Zika virus and fetopathy: a prospective cohort study
in French Guiana. Ultrasound Obstet Gynecol. 2017;49:729–36. https://
doi.org/10.1002/uog.17404.
Counotte MJ, Egli‑Gany D, Riesen M, Abraha M, Porgo TV, Wang J, et al.
Zika virus infection as a cause of congenital brain abnormalities and
Guillain‑Barré syndrome: from systematic review to living systematic
review. F1000Res. 2018;7:196. https://doi.org/10.12688/f1000research.
13704.1.
Freitas DA, Souza‑Santos R, Carvalho LMA, Barros WB, Neves LM, Brasil
P, et al. Congenital Zika syndrome: a systematic review. PLoS ONE.
2020;15:e0242367–e0242367. https://doi.org/10.1371/journal.pone.
0242367.
Costello A, Dua T, Duran P, Gülmezoglu M, Oladapo OT, Perea W, et al.
Defining the syndrome associated with congenital Zika virus infection.
Bull World Health Organ. 2016;94:406‑406A. https://doi.org/10.2471/
BLT.16.176990.
Romer Y, Valadez‑Gonzalez N, Contreras‑Capetillo S, Manrique‑Saide
P, Vazquez‑Prokopec G, Pavia‑Ruz N. Zika virus infection in pregnant
women, Yucatan. Mexico Emerg Infect Dis. 2019;25:1452–60. https://
doi.org/10.3201/eid2508.180915.
Brasil P, Pereira JP, Moreira ME, Ribeiro Nogueira RM, Damasceno L,
Wakimoto M, et al. Zika virus infection in pregnant women in Rio de
Janeiro. N Engl J Med. 2016;375:2321–34. https://doi.org/10.1056/
NEJMoa1602412.
Hill SC, Vasconcelos J, Neto Z, Jandondo D, Zé‑Zé L, Aguiar RS, et al.
Emergence of the Asian lineage of Zika virus in Angola: an outbreak
investigation. Lancet Infect Dis. 2019;19:1138–47. https://doi.org/10.
1016/S1473‑3099(19)30293‑2.
Lourenço J, de Lourdes Monteiro M, Valdez T, Monteiro Rodrigues J,
Pybus O, Rodrigues Faria N. Epidemiology of the Zika virus outbreak in
the Cabo Verde Islands, West Africa. PLoS Curr. 2018;10. doi:https://doi.
org/10.1371/currents.outbreaks.19433b1e4d007451c691f138e1e67e8c
Weetman D, Kamgang B, Badolo A, Moyes CL, Shearer FM, Coulibaly M,
et al. Aedes mosquitoes and Aedes‑borne arboviruses in Africa: current
and future threats. Int J Environ Res Public Health. 2018;15:220. https://
doi.org/10.3390/ijerph15020220.
Zaganjor I, Sekkarie A, Tsang BL, Williams J, Razzaghi H, Mulinare J, et al.
Describing the prevalence of neural tube defects worldwide: a system‑
atic literature review. PLoS ONE. 2016;11:e0151586. https://doi.org/10.
1371/journal.pone.0151586.
Chepkorir E, Tchouassi DP, Konongoi SL, Lutomiah J, Tigoi C, Irura Z,
et al. Serological evidence of Flavivirus circulation in human popula‑
tions in Northern Kenya: an assessment of disease risk 2016–2017. Virol
J. 2019;16:65. https://doi.org/10.1186/s12985‑019‑1176‑y.
Hunsperger E, Odhiambo D, Makio A, Alando M, Ochieng M, Omballa
V, et al. Zika virus detection with 2013 serosurvey, Mombasa. Kenya
Emerg Infect Dis J. 2020;26:1603. https://doi.org/10.3201/eid2607.
191363.
Rathore APS, St John AL. Cross‑reactive immunity among flaviviruses.
Front Immunol. 2020;11:334 Available: (https://www.frontiersin.org/
article/10.3389/fimmu.2020.00334).
Aubry F, Jacobs S, Darmuzey M, Lequime S, Delang L, Fontaine A, et al.
Recent African strains of Zika virus display higher transmissibility and
fetal pathogenicity than Asian strains. Nat Commun. 2021;12:916.
https://doi.org/10.1038/s41467‑021‑21199‑z.
Samy AM, Thomas SM, El WAA, Cohoon KP, Townsend PA. Mapping the
global geographic potential of Zika virus spread. Mem Inst Oswaldo
Cruz. 2016;111(9):559–60. https://doi.org/10.1590/0074‑02760160149.
Harris PA, Taylor R, Thielke R, Payne J, Gonzalez N, Conde JG. Research
electronic data capture (REDCap)‑a metadata‑driven methodology
and workflow process for providing translational research informatics
support. J Biomed Inform. 2009;42(2):377–81. https://doi.org/10.1016/j.
jbi.2008.08.010.
Villar J, Ismail LC, Victora CG, Ohuma EO, Bertino E, Altman DG, et al.
International standards for newborn weight, length, and head cir‑
cumference by gestational age and sex: the Newborn Cross‑Sectional
Study of the INTERGROWTH‑21st Project. Lancet. 2014;384:857–68.
https://doi.org/10.1016/S0140‑6736(14)60932‑6.
Centers for Disease Control and Prevention. Zika MAC‑ELISA: instruc‑
tions for use. Atlanta; 2018. Available: https://www.fda.gov/medic
Page 10 of 11
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
36.
37.
38.
al‑ devices/emergency‑situations‑medical‑ devices/emergency‑use‑
authorizations‑medical‑ devices#zika
Roehrig JT, Hombach J, Barrett ADT. Guidelines for plaque‑reduction
neutralization testing of human antibodies to dengue viruses. Viral
Immunol. 2008;21:123–32. https://doi.org/10.1089/vim.2008.0007.
Nguyen CT, Moi ML, Le TQM, Nguyen TTT, Vu TBH, Nguyen HT, et al.
Prevalence of Zika virus neutralizing antibodies in healthy adults in
Vietnam during and after the Zika virus epidemic season: a longitudi‑
nal population‑based survey. BMC Infect Dis. 2020;20:332. https://doi.
org/10.1186/s12879‑020‑05042‑2.
World Health Organization. Zika virus disease: interim case defini‑
tion,12 February 2016. 2016. Available: http://www.who.int/csr/disea
se/zika/case‑ definition/en/.
Marchi S, Viviani S, Montomoli E, Tang Y, Boccuto A, Vicenti I, et al. Zika
virus in West Africa: a seroepidemiological study between 2007 and
2012. Viruses. 2020;12:641. https://doi.org/10.3390/v12060641.
R Core Team. R Development Core Team. R: a language and environ‑
ment for statistical computing. 2017. pp. 275–286. http://www.R‑proje
ct.org
Langat SK, Eyase FL, Berry IM, Nyunja A, Bulimo W, Owaka S, et al. Ori‑
gin and evolution of dengue virus type 2 causing outbreaks in Kenya:
evidence of circulation of two cosmopolitan genotype lineages. Virus
Evol. 2020;6(1):veaa026. https://doi.org/10.1093/ve/veaa026.
Konongoi L, Ofula V, Nyunja A, Owaka S, Koka H, Makio A, et al.
Detection of dengue virus serotypes 1, 2 and 3 in selected regions
of Kenya: 2011–2014. Virol J. 2016;13:182. https://doi.org/10.1186/
s12985‑016‑0641‑0.
Barsosio HC, Gitonga JN, Karanja HK, Nyamwaya DK, Omuoyo DO,
Kamau E, et al. Congenital microcephaly unrelated to flavivirus expo‑
sure in coastal Kenya. Wellcome open Res. 2019;4:179. https://doi.org/
10.12688/wellcomeopenres.15568.1.
Orioli IM, Dolk H, Lopez‑Camelo JS, Mattos D, Poletta FA, Dutra
MG, et al. Prevalence and clinical profile of microcephaly in South
America pre‑Zika, 2005–14: prevalence and case‑control study. BMJ.
2017;359:j5018. https://doi.org/10.1136/bmj.j5018.
Morris JK, Rankin J, Garne E, Loane M, Greenlees R, Addor M‑C, et al.
Prevalence of microcephaly in Europe: population based study. BMJ.
2016;354:i4721. https://doi.org/10.1136/bmj.i4721.
Shen S, Xiao W, Zhang L, Lu J, Funk A, He J, et al. Prevalence of congeni‑
tal microcephaly and its risk factors in an area at risk of Zika outbreaks.
BMC Pregnancy Childbirth. 2021;21:214. https://doi.org/10.1186/
s12884‑021‑03705‑9.
Gobillot TA, Kikawa C, Lehman DA, Kinuthia J, Drake AL, Jaoko W, et al.
Zika virus circulates at low levels in western and coastal Kenya. J Infect
Dis. 2020;222:847–52. https://doi.org/10.1093/infdis/jiaa158.
Kayiwa JT, Nankya AM, Ataliba IJ, Mossel EC, Crabtree MB, Lutwama JJ.
Confirmation of Zika virus infection through hospital‑based sentinel
surveillance of acute febrile illness in Uganda, 2014–2017. J Gen Virol.
2018;99:1248–52. https://doi.org/10.1099/jgv.0.001113.
Duong V, Ong S, Leang R, Huy R, Ly S, Mounier U, et al. Low circulation
of Zika virus, Cambodia, 2007–2016. Emerg Infect Dis. 2017;23:296–9.
https://doi.org/10.3201/eid2302.161432.
Willcox AC, Collins MH, Jadi R, Keeler C, Parr JB, Mumba D, et al.
Seroepidemiology of dengue, Zika, and yellow fever viruses among
children in the Democratic Republic of the Congo. Am J Trop Med Hyg.
2018;99:756–63. https://doi.org/10.4269/ajtmh.18‑0156.
Borchering RK, Huang AT, Mier‑y‑Teran‑Romero L, Rojas DP, Rodri‑
guez‑Barraquer I, Katzelnick LC, et al. Impacts of Zika emergence
in Latin America on endemic dengue transmission. Nat Commun.
2019;10:5730. https://doi.org/10.1038/s41467‑019‑13628‑x.
Harris M, Caldwell JM, Mordecai EA. Climate drives spatial vari‑
ation in Zika epidemics in Latin America. Proceedings Biol Sci.
2019;286:20191578. https://doi.org/10.1098/rspb.2019.1578.
Aubry F, Dabo S, Manet C, Filipović I, Rose NH, Miot EF, et al. Enhanced
Zika virus susceptibility of globally invasive Aedes aegypti populations.
Science. 2020;370:991–6. https://doi.org/10.1126/science.abd3663.
Paixão ES, Teixeira MG, Costa M da CN, Barreto ML, Rodrigues LC.
Symptomatic dengue during pregnancy and congenital neurologic
malformations. Emerg Infect Dis. 2018;24:1748–50. https://doi.org/10.
3201/eid2409.170361.
Osoro et al. BMC Medicine
(2022) 20:291
Page 11 of 11
39. Marinho PS, Cunha AJ, Amim Junior J, Prata‑Barbosa A. A review of
selected Arboviruses during pregnancy. Matern Heal Neonatol Perina‑
tol. 2017;3:17. https://doi.org/10.1186/s40748‑017‑0054‑0.
40. Kariyawasam S, Senanayake H. Dengue infections during pregnancy:
case series from a tertiary care hospital in Sri Lanka. J Infect Dev Ctries.
2010;4:767–75. https://doi.org/10.3855/jidc.908.
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