PMLS REVIEWER

Clinical Laboratory – a facility subdivided into different

sections where common diagnostic procedures are done
by specialized health professionals.
The Clinical Laboratory Law
• Regulated by the Bureau of Health Facilities and
Republic Act No. 4688 (The Clinical Laboratory Law of Services under Department of Health (DOH).
1966)
o To prevent the operation of substandard,
• An act regulating the operation and maintenance of improperly managed, and poorly
clinical laboratories and requiring the registration equipped clinical laboratories.
of the same with the department of health,
providing penalty for violation thereof, and for other
purposes.
Based on Ownership:

• It was approved on June 18, 1966, by President 1. Government – operated and maintained by a
Ferdinand Edralin Marcos. government unit.

2. Private – owned, operated, and established by any

SECTION 1, states that: individual, corporation, association, or
Any person, firm, or corporation, operating and maintaining organization.
a clinical laboratory in which body fluids, tissues,
secretions, excretions and radioactivity from beings or Based on Function:
animals are analyzed for the determination of the presence
of pathologic organisms, processes and/or conditions in 1. Clinical Pathology
the persons or animals from which they were obtained, o Clinical Chemistry
shall register and secure a license annually at the office of o Hematology
the Secretary of Health: provided, that government hospital o Immunohematology
laboratories doing routine or minimum laboratory o Serology
examinations shall be exempt from the provisions of this o Parasitology
sections if their services are extensions of government o Bacteriology
regional or central laboratories. o Mycology
o Clinical Microscopy
o Toxicology
SECTION 2, states that: o Therapeutic Drug Monitoring
o Blood Banking
It shall be unlawful for any person to be professionally in- o Lab Endocrinology
charge of a registered clinical laboratory unless he is a o And other similar disciplines
licensed physician duly qualified in laboratory medicine
and authorized by the Secretary of Health, such 2. Anatomic Pathology
authorization to be renewed annually. No license shall be o Surgical Pathology
granted or renewed by the Secretary of Health for the o Immunohistopathology
operation and maintenance of a clinical laboratory unless o Cytology
such laboratory is under the administration, direction, and o Autopsy
supervision of an authorized physician, as provided for in o Forensic Pathology
the preceding paragraph. o Molecular Pathology

SECTION 3, states that: Based on Institutional Character:

The Secretary of Health, through the Bureau of Research 1. Institution-based – laboratory that operates within
and Laboratories shall be charged with the responsibility of the premises and is a part of an institution.
strictly enforcing the provisions of this Act and shall be
authorized to issue such rules and regulations as may be 2. Freestanding – laboratory that does not form part
necessary to carry out its provisions. of any other institution.

Based on Service Capability:

SECTION 4, states that:
1. Primary – has a minimum space requirement of 10
Any person, firm, or corporation who violates any provisions
square meters.
of this Act or the rules and regulations issued thereunder by
o Routine Hematology
the Secretary of Health shall be punished with
o Routine Urinalysis
imprisonment for not less than one month but not more
o Routine Fecalysis
than one year, or by a fine of not less than one thousand
o Qualitative Platelet
pesos nor more than five thousand pesos, or both such fine
o Blood typing (hospital-based)
and imprisonment, at the discretion of the court.
 2. Secondary – has a minimum space requirement of 5. San Lazaro Hospital/STD-AIDS Central
20 square meters. Cooperative Laboratory
o All tests in primary laboratory • HIV/AIDS and other sexually-transmitted
o Routine Chemistry infections
o Quantitative Platelet
o Cross-matching
o Gram staining Satellite Testing Site – testing site that performs laboratory
o KOH (hospital-based) examinations under the control of a licensed laboratory
outside the physical confines of the laboratory.
3. Tertiary – has a minimum space requirement of 60
square meters.
o All tests in secondary laboratory
Mobile Clinical Laboratory – testing unit that moves from
o Special Chemistry
one testing site to another.
o Special Hematology
o Immunology-Serology ❖ Must have a base laboratory.
o Microbiology
❖ Must collect specimens only.

Limited-Service Capability
❖ Must operate only within a 100-kilometre radius
1. Dialysis centers from its base laboratory.
2. Social hygiene clinics
Sections/Departments:

Special Clinical Laboratories – laboratories that provide • Hematology

highly specialized lab services that are not provided by a • Immuno-hematology (BB)
general clinical lab. • Immuno-serology
• Clinical Microscopy (Uri-Para)
• Clinical Chemistry
National Reference Laboratory (NRL) – laboratory in a • Microbiology (Bacteriology)
government hospital which is designated to provide special • Histo-pathology
functions and services such as:

• Confirmatory testing
• Surveillance Hematology
• Resolution of conflicts
Hematology – scientific study of blood and its components
• Training and research
that helps physicians diagnose abnormalities in the blood.
• Evaluation of kits and reagents
• External quality assessment program

Phlebotomy – standard procedure of blood collection using

lancets and needles of varying gauges.
NRLs in the Philippines
1. Skin puncture – capillaries (for kids)
1. National Kidney and Transplant Institute
• Hematology 2. Venipuncture – veins (for adults)
• Immunohematology
• Automated Urinalysis 3. Arterial puncture – arteries
• Immunopathology

2. East Avenue Medical Center Whole blood – composed of plasma (least dense
• Toxicology component), buffy coat and erythrocytes (most dense
component).
3. Lung Center of the Philippines
• Clinical Chemistry • 55% plasma
o 91.5% H2O
4. Research Institute for Tropical Medicine o Proteins 7.5%
• Tuberculosis ▪ 54% Albumins – smallest, most
• Mycology abundant transport fatty acids
• Transfusion-transmissible infections ▪ 38% Globulins – from plasma and
• Bacterial Diseases liver cells
• Antimicrobial Resistance ▪ 7% Fibrinogen + trace substances
o 1% Solutes/waste
• Influenza
• Malaria
• 45% formed elements
• Measles
o 99% RBCs
• And other exanthems, Rotavirus, Polio
o 1% WBCs and platelets
 ▪ 65% Neutrophil 3. Eosinophil – usually with a bi-lobed nucleus and
▪ 23% Lymphocyte has granules that are stained bright reddish orange
▪ 5% Monocyte that act as the defense against parasites and
▪ 4% Eosinophil activate our allergic response.
▪ 1% Basophil

Plasma – pale yellow liquid (intravascular/extracellular)

that transports RBCs, WBCs and platelets through the
blood vessels and removes waste products of metabolism.

• Water, sugar, fat, protein, and salt solution

• 55% of a normal human’s blood volume

4. Basophil – with purple-blue granules that are for

Red blood cell (erythrocyte) – anucleated cells produced the inflammatory response and are involved in the
from the bone marrow that transport oxygen from the lungs allergic response.
to the tissues and transport carbon dioxide back to the
lungs.

• Has hemoglobin – gas transporting protein

molecule that gives blood the red color it has.

• Associated conditions:
o Anemia – low number of RBCs

o Polycythemia – high number of RBCs

5. Monocyte – kidney-shaped or horseshoe-shaped
nucleus.
White blood cell (leukocyte) – nucleated cells that lacks • Dendritic cell: marks out cells that are
hemoglobin and acts as defense against infection. antigens (foreign bodies) that should be
1. Neutrophil – the most numerous in number that destroyed by lymphocytes
has a multi-lobed nucleus and is made up of pale
lilac granules, mainly for immune defense. • Macrophage – act as antigen-presenting
cells that are larger and lives longer than
a. Immune defense: protect the body from neutrophils
infection by killing and ingesting bacteria,
fungi, and foreign bodies. o They will present antigens by
digesting the pathogen and place
a remnant of the antigen on their
surface and present it to
lymphocytes for recognition.

2. Lymphocyte – has a spherical nucleus and a

“robin’s egg blue” cytoplasm.

a. T-cell – cellular immune response which

recognize foreign substances and process
them for removal
Platelets (thrombocytes) – cell fragments that form clots
b. B cell – antibody production during injury to prevent blood from leaking out.

c. NK cells – kills cancer cells

Tests Performed in Hematology Section

Hematology Section:

• Sample: whole blood and blood films

• Complete Blood Count (CBC)
o Hemoglobin
o Hematocrit
o Red blood cell count
 o White blood cell count Blood Cell Count
o Platelet count/estimate
a) Manual
o RBC indices (MCV, MCH, MCHC)
▪ MCV - Mean Corpuscular Volume
▪ MCH - Mean Corpuscular
Hemoglobin
▪ MCHC - Mean Corpuscular
Hemoglobin Concentration

Procedures performed in the Hema Section:

1. Counting the number or concentration of cells

2. Determining the relative distribution of various
types of cells
3. Measuring biochemical abnormalities of the blood
4. Hemostasis and coagulation assays
b) Automated

Hemoglobin Determination 1. Electrical Impedance – also known as Coulter

Principle where sizing and counting of particles
Hemoglobin – iron-containing oxygen transport
metalloprotein in the red blood cells. is based on changes in electrical resistance
creating voltage pulses.
Methods:
2. Optical Detection – Hydrodynamic focusing
a) Cyanmethemoglobin Method
method where it uses a laser light in cell
o Reference Method
counting and sizing.
o Reagent: Drabkin’s Reagent
o Principle: oxidation of ferrous iron to ferric
by potassium ferricyanide = Peripheral Blood Smear
methemoglobin then it is converted to
cyanmethemoglobin with cyanide ions 1. Place a small drop of blood on a VERY CLEAN slide.
Hold a second slide at the angle shown.
o Instrument: Spectrophotometer (540 nm)

b) Automated Hemoglobinometry – utilizes 2. While maintaining contact with the bottom slide,
cyanmethemoglobin method with modified pull the top slide back to contact the drop, which
Drabkin’s reagent. will spread by capillary action.

3. Maintain firm contact with the bottom slide and

c) Point-of-care (POC) Hemoglobin Assay –
HemoCue method that uses a modified push the top slide in one motion to produce the
smear.
azidemethemoglobin reaction with a reagent of
sodium nitrate and sodium azide.

Hematocrit Determination

Hematocrit – known as packed cell volume (PCV) or

erythrocyte volume fraction (EVF) which is for evaluation or
treatment of anemia and determines the presence of
nutritional deficiencies.

Methods:

a) Spun microhematocrit
o Manual procedure
o Blood collection method: skin puncture
o Spin a blood-filled capillary tube using a Reticulocyte Count
microhematocrit centrifuge Reticulocyte – young RBCs without nucleus but still bears
cytoplasmic RNA that determines how the bone marrow
b) Automated – computed from the mean cell produce and release new RBCs to compensate
volume and the red cell count. lost/damaged RBCs. The blood film is stained with
supravital stain.
Erythrocyte Sedimentation Rate - rate at which RBCs fall
in a column in known as erythrocyte sedimentation rate.

o Non-specific test for inflammation

o Reference Method: Westergren Method
o Anticoagulant: Sodium Citrate
o Time of testing: One hour

Plasma Coagulation Assays

4. John Needham – believed that there must be a “life
Prothrombine Time (PT) and Activated Partial source” that causes inanimate matter to
Thromboplastin Time (APTT) spontaneously come to life.

o Coagulation Testing
o Detect abnormalities in hemostasis
o Anticoagulant: Sodium Citrate
o Sample: Plasma only

Microbiology 5. Lazzaro Spallanzani – he observed that microbes

move through the air as possible source of
Microbiology – study of organisms too small to be seen by
contamination and can be destroyed by boiling.
the unaided eye.

1. Clinical Microbiology – study of microbial

pathogens considered health threats to people.

2. Diagnostic Microbiology – examination and

identification of organisms through laboratory
tests.
6. Louis Pasteur – developed the principles of
3. Food Microbiology – practical application and use vaccination, microbial fermentation, and
of beneficial microorganisms in food processing. pasteurization.

7. Joseph Lister – pioneer of antiseptic surgery and

Branches of Microbiology
introduced the use of carbolic acid (phenol) as a
1. Parasitology – study of parasites chemical sterilizing agent for surgical instruments.

2. Mycology – study of fungi 8. Hans Christian Gram – credited for the Gram
staining technique which distinguishes two major
3. Bacteriology – study of bacteria groups of bacteria: Gram-positive and Gram-
negative.
4. Virology – study of viruses
9. Alexander Fleming – discovered the first
antibiotic, Penicillin G, from a mold Penicillium
Pioneers in Microbiology notatum.

1. Girolamo Fracastoro – believed that diseases are 10. Robert Koch – established the theory of etiologic
caused by different types of rapidly multiplying agents cause diseases by providing experimental
minute body and that these bodies are transferred steps (Koch’s postulates) used to prove that a
from the infector to the infected in three ways: specific microbe causes a specific disease.
o By direct contact;
o By carriers such as soiled clothing and o Koch’s postulates – 4 generalized
linen; principles linking specific microorganisms
o Through the air. to specific diseases that remain today as
the “gold standard” in medical
2. Anton Van Leeuwenhoek – father of bacteriology microbiology.
and protozoology that discovered many life forms
called “animalcules”. He also made a single lens
microscope which enable the study of minute
organisms.

3. Francesco Redi – he disputed the theory of

spontaneous generation and performed an
experiment on decaying meat in 1668.
Bacterial Cell o Sulfur granules: Nocardia and Actinomyces
species
o Bipolar bodies: Yersinia pestis

Spores – structures that allow the bacteria to resist

sterilization that is composed of calcium dipicolinate.

o Terminal: Clostridium tetani

o Subterminal: Clostridium botulinum
o Central: Bacillus anthracis

Bacterial Forms

Spheres (Cocci)

o Diplococci (Streptococcus pneumoniae)

o Streptococci (Streptococcus pyogenes)
o Tetrad
o Staphylococci (Staphylococcus aureus)
Cell Membrane – the lipoprotein layer that surrounds the o Sarcina (Sarcina ventriculi)
cytoplasm and regulates the transport of solutes in and out
of the cell.

Rods (Bacilli)

Cell Wall – the semi-rigid casing that provides structural o Chain of bacilli (Bacillus anthracis)
shape and support to the cell. o Flagellate rods (Salmonella typhi)
o Spore-former (Clostridium botulinum)

Ribosomes – the site of protein synthesis and gives

granular structure to the cytoplasm. Spirals

o Vibrios (Vibrio cholerae)

o Spirilla (Helicobacter pylori)
Nucleoid – the region where the DNA is concentrated. o Spirochaetes (Treponema pallidum)

Capsule – the protective layer of a bacterium that resist Common Bacterial Pathogens
phagocytosis and desiccation.
BACTERIAL SPECIES DISEASES
Bacillus anthracis Anthrax
Pili – hair-like proteinaceous structures that extend from
the cell membrane into the external environment. Clostridium botulinum Botulism/food poisoning
o Neisseria gonorrheae has two types: somatic pili Corynebacterium
Diphtheria
for adhesion and sex pili for conjugation. diphtheriae
Escherichia coli Urinary tract infection

Flagellum – the structure that allows the bacteria to move. Staphylococcus aureus Pyogenic infections

o Atrichous – absence of flagellum. Streptococcus pyogenes Strep throat, scarlet fever

o Monotrichous – one polar flagellum Salmonella typhi Typhoid fever

o Amphitrichous – single flagellum on both ends Streptococcus

Pneumonia
pneumoniae
o Lophotrichous – tuft of flagella on either end or Treponema pallidum
Syphilis
subsp. pallidum
both ends
Mycobacterium
Tuberculosis
tuberculosis
o Peritrichous – flagella all around the organism
Helicobacter pylori Gastric ulcer

Inclusion Bodies – food reserves of the bacteria

o Babes-Ernst bodies: Corynebacterium

diphtheriae
o Much’s granules: Mycobacterium tuberculosis
Bacterial Metabolism and Growth Gram Stain Procedure

o Oxygen 1. Flood with crystal violet for 1 minute. Rinse.

o Aerobes – require oxygen for growth 2. Flood with iodine for 1 minute. Rinse
(obligate, facultative, microaerophilic) 3. Decolorize with alcohol until no more purple
o Anaerobes – grow best in an atmosphere comes off the slide. Rinse.
of reduced oxygen 4. Flood with safranin. Rinse and dry.

o Carbon Dioxide
o Capnophiles: need 5-10% carbon dioxide
to live
o Placed in candle jars

o Nutrients
o Autrotrophs – able to make energy-
containing organic molecules from
inorganic raw material by using basic
energy sources such as sunlight.
o Heterotrophs – organisms that must
make use of food that comes from other
organisms in the form of fats,
carbohydrates, and proteins.

o Temperature Acid-Fast Staining

o Psychrophile/cryophile: 0-15 °C
o Mesophile: 20-45 °C
o Thermophile: 50-60 °C
o Hyperthermophile: 8-113 °C

o Hydrogen and Ion Concentration (pH)

o Acidophile: pH 0-5.5 (Sulfolobus)
o Neutrophile: pH 5.5-8.0 (E.coli)
o Alkalophile: pH 8.5-11.5 (Vibrio cholerae)

Bacterial Growth Curve

Antibiotics – drugs administered to either kill bacteria or
1. Lag phase: no increase in number of living inhibit their growth by preventing reproduction.
bacterial cells.
o Bacteriostatic – agents that inhibit bacterial
2. Log phase: exponential increase in number of growth.
living bacterial cells.
o Bactericidal – agents that actively kill bacteria.
3. Stationary phase: plateau in number of living
bacterial cells; rate of cell division and death Mechanisms of Action
roughly equal.
o Inhibits cell wall, protein, and nucleic acid
4. Death or decline phase: exponential decreases in synthesis.
number of living bacterial. o Cell membrane destruction

Bacterial Staining
Antibiotic Susceptibility Testing – for therapeutic
1. Simple Stain guidelines that helps indicate which antibiotic is effective in
a. One stain is used (e.g., methylene blue) killing the bacteria causing the infection or disease.
b. Organisms should only be observed for
o Disk diffusion susceptibility test/Kirby-Bauer
size, shape, and uniformity of staining.
Method
o Broth dilution susceptibility test
2. Differential Stain
a. Used to distinguish between groups of
bacteria.
b. Gram staining, acid-fast staining Broth Dilution

1. Minimal inhibitory concentration – lowest

concentration of antimicrobial agent that inhibits
bacterial growth (bacteriostatic).
 2. Minimal bactericidal concentration – lowest
antibiotic concentration that results in 99.9%
death of the bacterial population (bactericidal).

Disk Diffusion Method – determines the susceptibility of

bacterial pathogens to antimicrobial agents where it based
on growth inhibition surrounding antibiotic impregnated
disks and measurement of the diameter of the zone of
inhibition.