Toxoplasmosis must also be considered in the differential diagnosis in any

immunosuppressed patient who has clinical or laboratory evidence of damage to the

central nervous system . 5

AccuDiag™ The organism is one of the most common latent infectious agents of man through out
the world. 6
Toxoplasma gondii IgG (Toxo IgG)
The Diagnostic Automation, Inc. Toxoplasma gondii IgG ELISA kit provides all the
ELISA Kit necessary reagents for the rapid quantitation of Toxoplasma gondiiIgG antibody in
human sera.

Cat# 1101-1 The sensitivity, specificity, and reproducibility of enzyme-linked immunosorbent

assays is comparable to other serological tests for antibody, such as
immunofluorescence, complement fixation, hemagglutination and radioimmunoassay.
11, 12, 13

96 Tests TEST PRINCIPLE

Enzyme-Linked Immunosorbent Assays (ELISA) rely on the ability of biological
Toxoplasma Gondii IgG materials (i.e., antigens) to adsorb to plastic surfaces such as polystyrene (solid phase).
Test When antigens bound to the solid phase are brought into contact with a patient's
ELISA
Enzyme Linked serum, antigen specific antibody, if present, will bind to the antigen on the solid phase
Method forming antigen-antibody complexes. Excess antibody is removed by washing. This is
Immunosorbent Assay
followed by the addition of goat anti -human IgG conjugated with horseradish
Indirect ELISA: Antigen peroxidase which then binds to the antibody-antigen complexes. The excess conjugate
Principle
Coated Plate is removed by washing, followed by the addition of Chromogen/Substrate,
Quantitative: Positive & tetramethylbenzidine (TMB). If specific antibody to the antigen is present in the
Detection Range
Negative Control patient's serum, a blue color develops. When the enzymatic reaction is stopped with
Sample 10 µL serum 1N H2SO4, the contents of the wells turn yellow. The color, which is indicative of the
concentration of antibody in the serum, can be read on a suitable spectrophotometer or
Total Time ~ 60 min.
ELISA microwell plate reader. 7, 8, 9, 10
12-14 Months from the
Shelf Life
manufacturing date
Specificity 100% SPECIMEN COLLECTION AND PREPARATION
Sensitivity 95.3% 1. Handle all blood and serum as if capable of transmitting infectious agents.
2. Optimal performance of the Diagnostic Automation, Inc. ELISA kit depends upon
the use of fresh serum samples (clear, non-hemolyzed, nonlipemic, non-icteric). A
INTENDED USE minimum volume of 50 µL is recommended, in case repeat testing is required.
The Diagnostic Automation/Cortez Diagnostics, Inc. Toxoplasma gondii (Toxo) IgG Specimens should be collected aseptically by venipuncture.18 Early separation
Enzyme-Linked Immunosorbent Assay (ELISA) is intended for the detection and from the clot prevents hemolysis of serum.
quantitative determination of IgG antibody to Toxoplasma gondii in human sera. This 3. Store serum between 2 and 8 °C if testing will take place within two days. If
product is not FDA cleared (approved) for use in testing (ie, screening) blood or specimens are to be kept for longer periods, store at -20° C or colder. Do not use a
plasma donors. For in vitro diagnostic Use. High complexity test. frost-free freezer because it may allow the specimens to go through freeze-thaw
cycles and degrade antibody. Samples that are improperly stored or are subjected
to multiple freeze-thaw cycles may yield erroneous results.
SUMMARY AND EXPLANATION 4. If paired sera are to be collected, acute samples should be collected as soon as
possible after the onset of symptoms. The second sample should be collected 14 to
Toxoplasma gondii is a coccidian parasite initially isolated in 1908 from a North 21 days after the acute specimen was collected. Both samples must be run in
African rodent the gondii. Since then, the organism has been found in many species of duplicate on the same plate to test for a significant rise. If the first specimen is
birds, reptiles and mammals. 1 obtained late during the course of the infection, a significant rise may not be
detectable.
Man is infected with Toxoplasma gondii from various suspected sources: 5. The NCCLS provides recommendations for storing blood specimens (Approved
ingestion of infected meat, especially mutton and pork, or ingestion of soil Standard - Procedures for the Handling and Processing of Blood Specimens, H18-
contaminated by oocysts from domestic and feral cat. 2 Transmission by organ A. 1990). 18
transplant, transfusion or activation of quiescent infections is also documented.
Congenital Toxoplamsosis is a disease with an extraordinarily wide range of
manifestations; so wide in fact, that it must be considered in the differential diagnosis MATERIALS AND COMPONENTS
of nearly all types of obscure illness occurring during infancy. 3
Materials provided with the test kits
Because symptoms are sometimes nonspecific (i.e., anemia, splenomegaly, jaundice, Each kit contains the following components in sufficient quantities to perform the
fever, hepatomegaly, adenopathy and vomiting), congenital Toxoplasmosis is easily number of tests indicated on the package label.
misdiagnosed on the clinical grounds, even in sick infants who have the generalized
form of the disease . 4 1. Toxoplasma gondii antigen (inactivated) coated microassay plate: 96 wells,
configured in twelve 1x8 strips, stored in a foil pouch with desiccant. (96T: one
plate; 480T: five plates)

DAI CODE #1 Page 1 of 6

2. Serum Diluent Type I: Ready for use. Contains proclin (0.1%) as a 1. Place the desired number of strips into a microwell frame. Allow four (4)
preservative (96T: one bottle, 30 mL, 480T: two bottles, 60 mL each) Control/Calibrator determinations (one Negative Control, two Calibrators
3. Calibrator: Human serum or defibrinated plasma. Sodium azide (< 0.1%) and and one Positive Control) per run. A reagent blank (RB) should be run on
pen/strep (0.01%) added as preservatives, with kit specific factor printed each assay. Check software and reader requirements for the correct
on vial label. The Calibrator is used to calibrate the assay to account for Control/Calibrator configuration. Return unused strips to the sealable bag
day-to-day fluctuations in temperature and other testing with desiccant, seal and immediately refrigerate.
conditions. (96T: one vial, 0.4 mL; 480T: one vial, 0.8 mL)
4. Positive Control: Human serum or defibrinated plasma. Sodium azide (< Example Configuration:
0.1%) and pen/strep (0.01%) added as preservatives, with established range
printed on vial label. The Positive Control is utilized to control the positive Plate Sample Plate Sample Description
range of the assay. (96T: one vial, 0.4 mL; 480T: one vial, 0.8 mL)* Location Description Location
5. Negative Control: Human serum or defibrinated plasma. Sodium azide (< 1A RB 2A Patient #4
0.1%) and pen/strep (0.01%) added as preservatives, with established
1B NC 2B Patient #5
range printed on vial label. The Negative Control is utilized to control the
1C Cal 2C Patient #6
negative range of the assay. (96T: one vial, 0.4 mL; 480T: one vial, 0.8 mL) *
1D Cal 2D Patient #7
6. Horseradish-peroxidase (HRP) Conjugate: Ready to use. Goat anti-human
1E PC 2E Patient #8 (Acute 1)
IgG, containing proclin (0.1%) and gentamicin as preservatives. (96T: one
1F Patient #1 2F Patient #8 (Acute 2)
bottle, 16 mL; 480T: five bottles, 16 mL each)
7. Chromogen/Substrate Solution Type I: Tetramethylbenzidine (TMB), ready 1G Patient #2 2G Patient #8 (Convalescent 1)
to use. The reagent should remain closed when not in use. If allowed to 1H Patient #3 2H Patient #8 (Convalescent 2)
evaporate, a precipitate may form in the reagent wells. (96T: one bottle, 15 mL;
480T: five bottles, 15 mL each) RB = Reagent Blank - Well without serum addition run with all reagents. Utilized to
8. Wash Buffer Type I (20X concentrate): Dilute 1 part concentrate + 19 parts blank reader.
deionized or distilled water. Contains TBS, Tween-20 and proclin (0.1%) as a NC = Negative Control
preservative. (96T: one bottle, 50 mL; 480T: one bottle, 250 mL) Cal = Calibrator
9. Stop Solution: Ready to use, contains a 1N H2SO4 solution. (96T: one bottle, PC = Positive Control
15 mL; 480T: five bottles, 15 mL each) 2. Dilute test sera, Calibrator and Control sera 1:21 (e.g., 10 µL + 200 µL) in
* Note: serum vials may contain excess volume Serum Diluent. Mix well. (For manual dilutions it is suggested to dispense
the Serum Diluent into the test tube first and then add the patient serum.)
3. To individual wells, add 100 µL of the appropriate diluted Calibrator,
Materials required but not provided Controls and patient sera. Add 100 µL of Serum Diluent to reagent blank
1. Wash bottle, automated or semi-automated microwell plate washing system. well. Check software and reader requirements for the correct reagent blank
2. Micropipettes, including multichannel, capable of accurately delivering 10-200 µL well configuration.
volumes (less than 3% CV). 4. Incubate each well at room temperature (21 to 25 °C) for 25 minutes +/- 5
3. One liter graduated cylinder. minutes.
4. Paper towels. 5. Aspirate or shake out liquid from all wells. If using semi-automated or
5. Test tube for serum dilution. automated washing equipment add 250-300 µL of diluted Wash Buffer to
6. Reagent reservoirs for multichannel pipettes. each well. Aspirate or shake out to remove all liquid. Repeat the wash
7. Pipette tips. procedure two times (for a total of three (3) washes) for manual or semi-
8. Distilled or deionized water (dh20), CAP (College of American Pathology) Type automated equipment or four times (for a total of five (5) washes) for
1or equivalent. 20, 21 automated equipment. After the final wash, blot the plate on paper toweling
9. Timer capable of measuring to an accuracy of +/- 1 second (0 - 60 minutes). to remove all liquid from the wells.
10. Disposal basins and 0.5% sodium hypochlorite (50 mL bleach in 950 mL dH20) **IMPORTANT NOTE: Regarding steps 5 and 8 - Insufficient or
11. Single or dual wavelength microplate reader with 450 nm filter. If dual excessive washing will result in assay variation and will affect validity of
wavelength is used set the reference filter to 600-650 nm. Read the Operator's results. Therefore, for best results the use of semi- automated or
Manual or contact the instrument manufacturer to establish linearity performance automated equipment set to deliver a volume to completely fill each well
specifications of the reader. (250-300 µL) is recommended. A total of up to five (5) washes may be
necessary with automated equipment. Complete removal of the Wash
Note: Use only clean, dry glassware. Buffer after the last wash is critical for the accurate performance of
the test. Also, visually ensure that no bubbles are remaining in the
METHODS FOR USE wells.
PREPARATION FOR THE ASSAY 6. Add 100 µL Conjugate to each well, including reagent blank well. Avoid
1. All reagents must be removed from refrigeration and allowed to come to room bubbles upon addition as they may yield erroneous results.
temperature before use (21° to 25° C). Return all reagents to refrigerator 7. Incubate each well at room temperature (21 to 25 °C) for 25 minutes +/- 5
promptly after use. minutes.
2. All samples and controls should be vortexed before use. 8. Repeat wash as described in Step 5.
3. Dilute 50 mL of the 20X Wash Buffer Type I to 1 L with distilled and/or 9. Add 100 µL Chromogen/Substrate Solution (TMB) to each well, including
deionized H20. Mix well. the reagent blank well, maintaining a constant rate of addition across the
plate.
10. Incubate each well at room temperature (21 to 25 °C) for 10-15 minutes.
11. Stop reaction by addition of 100 µL of Stop Solution (1N H2SO4) following
ASSAY PROCEDURE the same order of Chromogen/Substrate addition, including the reagent
blank well. Tap the plate gently along the outsides, to mix contents of the
Note: To evaluate paired sera, both serum samples must be tested in duplicate
wells. The plate may be held up to 1 hour after addition of the Stop
and run in the same plate. It is recommended that the serum pairs be run in
Solution before reading.
adjacent wells.

DAI CODE #1 Page 2 of 6

 12. The developed color should be read on an ELISA plate reader equipped Acute 1 ISR = 0.8 to 1.2 Convalescent 1 ISR= 0.8 to 1.2
with a 450 nm filter. If dual wavelength is used, set the reference filter to Acute 2 ISR Convalescent 2 ISR
600-650 nm. The instrument should be blanked on air.
The reagent blank must be less than 0.150 Absorbance at 450 nm. If the reagent blank
is > 0.150 the run must be repeated. Blank the reader on the reagent blank well and
then continue to read the entire plate. Dispose of used plates after readings have been 6. Compare the ISR of the pairs by calculating as follows:
obtained.
Mean ISR (second sample) - Mean ISR (first sample) X 100 = % RISE IN ISR LEVEL
Mean ISR (first sample)
RESULTS
1. Mean Calibrator O.D. (Optical Density) - Calculate the mean O.D. value INTERNATIONAL UNIT CONVERSION
from the two Calibrator determinations. International unit (IU) reactivity is determined relative to the IU standard. Conversion
2. Correction Factor - To account for day-to-day fluctuations in assay activity of Index values to international units is accomplished by using an exponential
due to room temperature and timing, a Correction Factor is determined by regression analysis. Each lot is standardized versus international units and provided
Diagnostic Automation/Cortez Diagnostics, Inc. for each lot of kits. The with a lot specific conversion table (Conversion of International Units (IU) per mL
Correction Factor is printed on the Calibrator vial. for Toxoplasma IgG). For example:
3. Cutoff Calibrator Value - The Cutoff Calibrator Value for each assay is
determined by multiplying the Correction Factor by the mean Calibrator ISR IU
O.D. determined in Step 1.
4. ISR Value - Calculate an Immune Status Ratio (ISR) for each specimen by 1.0 19.5
dividing the specimen O.D. Value by the Cutoff Calibrator Value 1.5 35
2.0 62
determined in Step 3.
2.5 111
Example: O.D.s obtained for Calibrator = 0.38, 0.42
Mean O.D. for Calibrator = 0.40 See attached addendum for the lot specific conversion table.
Correction Factor = 0.50
Cutoff Calibrator Value = 0.50 x 0.40 = 0.20 % Rise in ISR Interpretation
O.D. obtained for patient sera = 0.60 <30.0 % No significant change in antibody level. No evidence
ISR Value = 0.60/0.20 = 3.00 of recent infection. If active disease is still suspected, a
third sample should be collected and tested in the same
ANALYSIS assay as the first sample to look for a significant rise in
1. The patients’ ISR (Immune Status Ratio) values are interpreted as follows: antibody level.
ISR Results Interpretation
≤ 0.90 Negative No detectable antibody to Toxoplasma gondii by the ≥ 30.0% Statistically significant change in antibody level
ELISA test. detected. This identifies those persons who are
Such individuals are presumed to be uninfected with presumed to be experiencing recent or current episodes
Toxoplasma Gondii and to be susceptible to primary of Toxo infection (reactivation, reinfection or a
infection. primary infection where the acute specimen was
obtained too late to demonstrate seroconversion).
0.91-1.09 Equivocal Samples should be retested. See Number (3) below.
≥1.10 Positive Indicates presence of detectable antibody to Note: When evaluating paired sera, it should be determined if samples with
Toxoplasma gondii by the ELISA test. Indicative of high absorbance values are within linearity specifications of the spectrophotometer.
current or previous infection.The individual may be at Read the Operator's Manual or contact the instrument's manufacturer to obtain the
risk of transmitting Toxoplasma gondii infection, but is established linearity specifications of your spectrophotometer.
not necessarily currently contagious.

2. Samples that remain equivocal after repeat testing should be retested on an QUALITY CONTROL
alter nate method, e.g., immunofluorescence assay (IFA). If results remain
equivocal upon further testing, an additional sample should be taken. (See For the assay to be considered valid the following conditions must be met:
Limitation No. 4). 1. Calibrator and Controls must be run with each test run.
3. In the evaluation of paired sera, if the acute specimen is negative and the 2. Reagent blank (when read against air blank) must be < 0.150 Absorbance (A) at
convalescent specimen is positive, a seroconversion has taken place. This 450 nm.
indicates a significant change in antibody level and the patient is 3. Negative Control must be < 0.250 A at 450 nm (when read against reagent
undergoing a primary infection. blank).Each Calibrator must be > 0.250 A at 450 nm (when read against reagent
4. To evaluate paired sera for a significant change in antibody level or blank).
seroconversion, both samples must be tested in duplicate in the same assay. 4. Positive Control must be > 0.500 A at 450 nm (when read against reagent blank).
The mean ISR of both samples (acute and onvalescent) must be greater 5. The ISR(Immune Status Radio) Values for the Positive and Negative Control
than 1.00 to evaluate the paired sera for significant rise in antibody level. should be in their respective ranges printed on the vials. If the Control values are
5. Additional Quality Control for Paired Sera: (See NOTE under Assay not within their respective ranges, the test should be considered invalid and should
Procedure). As a check for acceptable reproducibility of both the acute be repeated.
sera (tested in duplicate) and the convalescent sera (tested in duplicate), 6. Additional Controls may be tested according to guidelines, or requirements of
the following criteria must be met for valid results: local, state, and/or federal regulations or accrediting organizations
7. Refer to NCCLS C24-A for guidance on appropriate QC practices. 19
8. If above criteria are not met upon repeat testing, contact Diagnostic Automation,
Inc. Technical Services.

DAI CODE #1 Page 3 of 6

 INTERNATIONAL UNIT CONVERSION
The data in Table 4 illustrate Toxoplasma IgG ISR Values for the serially diluted
PERFORMANCE CHARACTERISTICS international unit standard, obtained from the World Health Organization. The
Toxoplasma IgG ISR Values are compared to serial dilutions of the international unit
SENSITIVITY AND SPECIFICITY standard serum by linear regression (exponential regression analysis). The data
A total of 106 samples from a random population were tested by the Diagnostic indicate that international units can be determined from the ISR Value.
Automation, Inc. Toxoplasma gondii IgG ELISA kit and a commercially available
Toxoplasma gondii IFA kit. Complete agreement was found for 101 of the samples, of
which 63 (62%) were negative and 38 (38%) were positive. Five samples were found Table 4
negative by the Diagnostic Automation, Inc. ELISA and positive (1:16, 1:16, International Unit Conversion
1:32,1:32, 1:32) by IFA. This data indicates 95.3% sensitivity and 100% specificity International Unit Standard Units ISR Value
when the Diagnostic Automation, Inc. Toxoplasma gondii IgG ELISA is compared to / mL
the IFA technique. Upon further testing with an alternate method as referee, all five 500 3.8
samples were found to be negative by concensus. 250 3.2
125 2.6
Forty-seven of the 106 samples were also compared with another commercially 62.5 2.0
available Toxoplasma gondii IgG ELISA kit. Complete agreement was found for 31.3 1.4
twenty-two samples that were positive and twenty-four samples that were negative. 15.6 0.8
One sample tested positive on the Diagnostic Automation, Inc. ELISA kit and
negative on the other ELISA kit. After further testing, with the a commercially Linear regression compared ISR Values versus International Units.
available Toxoplasma gondii IgG IFA kit used as a referee method, the sample was
confirmed positive.
r2 = 0.995 a = 0.864 b = 1.566 Y = ISR X = IU / mL
Ten paired sera (twenty samples), were evaluated with the Diagnostic Automation,
Inc. Toxoplasma gondiiIgG ELISA kit, another commercially available ELISA kit Exponential Regression Equation Calculation
and a commercially available Tox oplasma gondii IFA kit. Each pair was taken
from an individual with diagnosed Toxoplasma gondii infection. Six of the X = (y+b) ex = derived IU / mL
individuals were positive for seroconversion (negative on acute and positive on a
convalescent) and four of the individuals were positive on both acute and convalescent
samples. All ten pairs showed a significant rise in antibody level. Complete
agreement, 100% sensitivity was established for the pairs when compared to the other
LIMITATION OF PROCEDURE
ELISA test and the IFA. 1. Use fresh serum samples or samples frozen only once and thawed at 37 °C.
Samples that are improperly stored or are subjected to multiple freeze-thaw
REPRODUCIBILITY cycles may yield spurious results.
Three studies were performed to assess the precision of the TMB test results. Five sera 2. The user of this kit is advised to carefully read and understand the package
were used in 20 wells each ranging from negative to high positive for the inter-run insert. Strict adherence to the protocol is necessary to obtain reliable test results.
assay. Five sera were used for 5 days in 5 wells each ranging from negative to high In particular, correct sample and reagent pipetting, along with careful washing
positive for the inter-day assay. Five sera were used in 3 wells each ranging from and timing of the incubation steps are essential for accurate results.
negative to high positive with 3 different lots of TMB Substrate. The summary of 3. This kit is designed to measure IgG antibody in patient samples. Positive results
results are as follows: in neonates must be interpreted with caution, since maternal IgG is transferred
Table 1 passively from the mother to the fetus before birth. IgM assays are generally
Intra-Run Assay more useful indicators of infection in children below 6 months of age.
4. Samples collected very early in the course of an infection may not have
Serum 1 Serum 2 Serum 3 Serum 4 Serum 5
detectable levels of IgG. In such cases, it is recommended that an IgM assay be
Mean = 1.425 4.275 3.239 0.318 0.182
performed, or a second serum sample be obtained at a later date to be tested in
S.D. = 0.107 0.087 0.187 0.023 0.016
parallel with the original sample to determine seroconversion. 15, 16
C.V. = 7.5% 2.0% 5.8% 7.2% 8.7%
5. The results of ELISA performed on serum from patients with
immunosuppression must be interpreted with caution. The presence of IgG
Table 2 antibody against a particular virus or organism may not assure protection from
Intra-Run Assay that disease. For example, cases of reactivation of Toxoplasma gondii infection
Serum 1 Serum 2 Serum 3 Serum 4 Serum 5 in immunocompromised individuals have been documented. 17 Alternatively,
Mean = 1.644 5.632 4.223 0.406 0.286 certain immune individuals have been shown to have such low circulating IgG
S.D. = 0.191 0.414 0.273 0.058 0.015 levels that they may appear negative for that antibody when tested. 6
C.V. = 11.6% 7.4% 6.5% 14.4% 5.4% 6. Samples that remain equivocal after repeat testing should be retested on an
alternate method, e. g., immunofluorescence assay (IFA). If results remain
equivocal upon further testing, an additional sample should be taken (See
Table 3 Limitation No. 4).
Intra-Run Assay 7. The values obtained from this assay are intended to be an aid to diagnosis only.
Serum 1 Serum 2 Serum 3 Serum 4 Serum 5 Each physician must interpret the results in light of the patient's history, physical
Mean = 1.81 6.40 4.565 0.39 0.266 findings and other diagnostic procedures.
S.D. = 0.22 0.347 0.322 0.063 0.062
C.V. = 12.1% 5.4% 7.0% 16.2% 23.3%
PRECAUTIONS
1. For in vitro diagnostic use.

DAI CODE #1 Page 4 of 6

2. The human serum components used in the preparation of the Controls and P501: Dispose of contents and container in accordance to local, regional, national and
Calibrator in this kit have been tested by an FDA approved method for the international regulations.
presence of antibodies to human immunodeficiency virus1 & 2 (HIV 1&2),
hepatitis C (HCV) as well as hepatitis B surface antigen and found negative. WARNING
Because no test method can offer complete assurance that HIV, HCV, hepatitis B Serum Diluent and Controls contain < 0.1% sodium azide.
virus, or other infectious agents are absent, specimens and human-based reagents H302: Harmful if swallowed
should be handled as if capable of transmitting infectious agents. P264: Wash thoroughly with plenty of soap and water after handling
3. The Centers for Disease Control & Prevention and the National Institutes of P270: Do not eat, drink or smoke when using this product
Health recommend that potentially infectious agents be handled at the Biosafety P301+P312: IF SWALLOWED: Call a POISON CENTER or doctor/physician if you
Level 2. 15 feel unwell
4. The components in this kit have been quality control tested as a Master Lot unit. P330: If swallowed, rinse mouth
Do not mix components from different lot numbers except Chromogen/Substrate P501: Dispose of contents/container to in accordance to local, regional, national and
Solution Type I, Stop Solution, Wash Buffer Type I. Do not mix with international regulations.
components from other manufacturers.
5. Do not use reagents beyond the stated expiration date marked on the package
label.
6. All reagents must be at room temperature (21° to 25° C) before running assay. STORAGE
Remove only the volume of reagents that is needed. Do not pour reagents back 1. Store unopened kit between 2° and 8° C. The test kit may be used throughout the
into vials as reagent contamination may occur. expiration date of the kit. Refer to the package label for the expiration date.
7. Before opening Control and Calibrator vials, tap firmly on the benchtop to 2. Unopened microassay plates must be stored between 2° and 8° C. Unused strips
ensure that all liquid is at the bottom of the vial. must be immediately resealed in a sealable bag with desiccant and returned to
8. Use only distilled or deionized water and clean glassware. storage between 2° and 8° C.
9. Do not let wells dry during assay; add reagents immediately after completing 3. Store HRP Conjugate between 2° and 8° C.
wash steps. 4. Store the Calibrator, Positive and Negative Controls between 2° and 8° C.
10. Avoid cross-contamination of reagents. Wash hands before and after handling 5. Store Serum Diluent Type I and 20X Wash Buffer Type 1 between 2° and 8° C.
reagents.Cross-contamination of reagents and/or samples could cause false 6. Store the Chromogen/Substrate Solutions Type 1 between 2° and 8° C. The
results. reagent should remain closed when not in use. If allowed to evaporate, a
11. If washing steps are performed manually, wells are to be washed three times. Up precipitate may form in the reagent wells.
to five wash cycles may be necessary if a washing manifold or automated 7. Store 1X (dilute) Wash Buffer Type 1 at room temperature (21° to 25° C) for up
equipment is used. to 5 days or up to 1 week between 2° and 8° C.
12. Sodium azide inhibits Conjugate activity. Clean pipette tips must be used
for the Conjugate addition so that sodium azide is not carried over from
other reagents. Note: If constant storage temperature is maintained, reagents and substrate will be
13. It has been reported that sodium azide may react with lead and copper in stable for the dating period of the kit. Refer to package label for expiration date.
plumbing to form explosive compounds. When disposing, flush drains with Precautions were taken in the manufacture of this product to protect the reagents from
water to minimize build-up of metal azide compounds. contamination and bacteriostatic agents have been added to the liquid reagents. Care
14. Never pipette by mouth or allow reagents or patient sample to come into contact should be exercised to protect the reagents in this kit from contamination.
with skin. Reagents containing ProClin®, sodium azide, and TMB may be
irritating. Avoid contact with skin and eyes. In case of contact, flush with plenty
of water. REFERENCES
15. If a sodium hypochlorite (bleach) solution is being used as a disinfectant, do not 1. Feldman, H. A. 1974. Toxoplasmosis: An Overview. Bull. N.Y. Acad. Med. 50:
expose to work area during actual test procedure because of potential 110-127.
interference with enzyme activity. 2. Jacobs, L. J. 1974. Toxoplasma gondii: Parasitology and Transmission. Bull. N.
16. Avoid contact of Stop Solution (1N sulfuric acid) with skin or eyes. If contact Y. Acad. Med. 50: 128- 145.
occurs, immediately flush area with water. 3. Eichenwald, H. F. 1959. A study of Congenital Toxoplasmosis with Particular
17. Caution: Liquid waste at acid pH must be neutralized prior to adding sodium Emphasis on, Sequelae and Therapy. In:Human Toxoplasmosis. Sim, J. C., ed.
hypochlorite (bleach) solution to avoid formation of poisonous gas. Recommend Vol. 2. Munksgaard, Copenhagen.
disposing of reacted, stopped plates in biohazard bags. See Precaution 3. 4. Alford, C. A., Jr., S. Stagno, and D. W. Reynolds. 1974. Congenital
18. The concentrations of anti-Toxoplasma gondii in a given specimen determined Toxoplasmosis: Laboratory, and Therapeutic Considerations, With Special
with assays from different manufacturers can vary due to differences in assay Reference to SubTrinity Biotechnical Disease. Bull. N. Y. Acad. Med. 50: 160-
methods and reagent specificity. 181.
5. Remington, J. S. 1974. Toxoplasmosis In The Adult. Bull. N. Y. Acad. Med.,
The safety data sheet is available upon request. 50:211-227.
6. Beattie, C. P. 1967. Toxoplasmosis. In: Recent Advances in Medical
Microbiology. A. P Waterson, ed. pp 318- 351.
7. Engvall, E., and P. Perlman. 1971. Enzyme-Linked Immunosorbent Assay,
(ELISA) Quantitative Assay of Immunoglobulin G. In: Immunochemistry. 8:
871-874.
WARNING 8. Engvall, E., and P. Perlman. 1971. Enzyme-Linked Immunosorbent Assay,
Serum Diluent, Conjugate, and Wash Buffer contain 0.1% ProClin 300R, a biocidal ELISA. Peeters. H., ed. In: Protides of the Biological Fluids. Proceedings of the
preservative that may cause sensitization by skin contact; prolonged or repeated Nineteenth Colloquium, Brugge, Oxford. Pergamon Press. pp 553-556.
exposure may cause allergic reaction in certain sensitive individuals. 9. Engvall, E., K. Jonsson, and P. Perlman.1971. Enzyme-Linked Immunosorbent
H317: May cause an allergic skin reaction. Assay. II. Quantitative Assay of Protein Antigen, Immunoglobulin-G, By
P280: Wear protective gloves / protective clothing / eye protection / face protection. Means of Enzyme-Labelled Antigen and Antibody-Coated tubes. In: Biochem.
P302 + P352: IF ON SKIN: Wash with plenty of soap and water. Biophys. Acta. 251: 427-434.
P333 + P313: If skin irritation or rash occurs: Get medical advice/ attention.

DAI CODE #1 Page 5 of 6

10. Van Weeman, B. K. and A.H.W.M. Schuurs. 1971. Immunoassay Using
Antigen-Enzyme Conjugates. In: FEBS Letter. 15: 232-235.
11. Walls, K. W. 1978. Serodiagnosis of Toxoplasmosis. Lab. Mgmt. January: 26-31
12. Bakerman, S. 1980. Enzyme Immunoassays. Lab. Mgmt. August: 21-29.
13. Engvall, E. and P. Perlman. 1972. Enzyme-linked Immunosorbent Assay,
ELISA. III. Quantitation of Specific Antibodies by Enzyme-Labeled Anti-
Immunoglobulins in Antigen Coated Tubes. J. Immunol. 109: 129-135.
14. CDC-NIH Interagency Working Group. 1993. In: Biosafety in Microbiological
and Biomedical Laboratories, 3rd Edition. U. S. Dept. of Health and Human
Services. Public Health Service. pp 9-12.
15. Christie, A. B. 1974. Toxoplasmosis. In: Infectious Diseases. 2nd Edition. p.
995-1007.
16. American Academy of Pediatrics. 1986. Toxoplasma gondii. In: Report of the
Committee on Infectious Diseases. G. Peters, ed. pp 366-369.
17. Shepp, D. H., et. al. 1985. Toxoplasma gondii. Reactivation Identified by
Detection of Parasitemia in Tissue Culture. Ann. Intern. Med. 103: 218-221.
18. National Committee for Clinical Laboratory Standards. 1990. Procedures for the
Collection of Diagnostic Blood Specimens by Venipuncture Approved Standard.
NCCLS Publication H18-A.
19. NCCLS. 1991. National Committee for Clinical Laboratory Standard. Internal
Quality Control Testing: Principles & Definition.NCCLS Publication C24-
20. http://www.cap.org/html/ftpdirectory/checklistftp.html. 1998. Laboratory
General - CAP (College of American Pathology) Checklist (April 1998). pp 28-
32.
21. NCCLS. 1997. National Committee for Clinical Laboratory Standard.
Preparation and Testing of Reagent Water in the Clinical Laboratory. NCCLS
Publication C3- A3.

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