MICROBIOLOGY

ANNUAL
#Laboratory Diagnosis of Hepatitis B Infection (5 marks):

1. Serology:

- HBsAg (Hepatitis B surface antigen): Detectable in acute and chronic infection; indicates active
infection.

- Anti-HBs (Hepatitis B surface antibody): Indicates immunity from past infection or vaccination.

- HBcAg (Hepatitis B core antigen): Not detected in blood but is present in infected hepatocytes.

- Anti-HBc (Hepatitis B core antibody): Appears in both acute and chronic infection, may indicate
prior exposure.

- HBeAg (Hepatitis B e antigen): Indicates high viral replication and infectivity.

- Anti-HBe (Hepatitis B e antibody): Suggests lower infectivity and viral replication in chronic
infection.

2. PCR:

- HBV DNA: Quantifies the viral load in the blood, useful in assessing viral replication and
monitoring antiviral therapy.

3. Liver Function Tests:

- Elevated liver enzymes (ALT, AST): Indicate hepatocellular injury during the acute phase.

- Elevated bilirubin: Seen in severe cases, indicating liver dysfunction.

4. Liver Biopsy:

- Used in chronic cases to assess the degree of liver damage, fibrosis, or cirrhosis.

3. Short Notes on the Following

a) Type IV Hypersensitivity (e.g., Tuberculosis) (2 marks):

- Definition: Type IV hypersensitivity is a delayed-type hypersensitivity (DTH) reaction mediated by T

lymphocytes rather than antibodies.
- Example: Tuberculin reaction in which CD4+ T cells recognize Mycobacterium tuberculosis antigens,
leading to inflammation and tissue damage.

- Mechanism: After antigen exposure, T cells release cytokines like TNF-α, which recruit
macrophages and cause tissue damage. This reaction peaks 48-72 hours after antigen exposure.

b) Bacterial Growth Curve (2 marks):

- The bacterial growth curve typically has four phases:

1. Lag Phase: Bacteria are adapting to the growth environment, no significant growth occurs.

2. Log Phase: Exponential growth, bacteria divide rapidly.

3. Stationary Phase: Nutrients become limited, and the rate of new cells formed equals the rate of
cell death.

4. Death Phase: Nutrient depletion and toxic accumulation cause bacteria to die at a greater rate
than they are being formed.

c) Oral Thrush (2 marks):

- Definition: Oral thrush is a fungal infection of the oral mucosa caused by *Candida albicans*,
characterized by white, curd-like plaques on the tongue, cheeks, and palate.

- Risk Factors: Immunocompromised states (e.g., HIV, chemotherapy), diabetes, antibiotic use,
dentures.

- Symptoms: Pain, difficulty swallowing, and a burning sensation in the mouth.

- Diagnosis: Direct microscopy with potassium hydroxide (KOH) preparation showing budding yeast
cells.

d) Autoclave (2 marks):

- Definition: An autoclave is a machine that uses high-pressure steam to sterilize equipment and
materials. It operates at temperatures of 121°C at 15 psi for 15-20 minutes.

- Applications: Used in laboratories, hospitals, and dental offices for sterilizing surgical instruments,
media, and laboratory glassware.

- Mechanism: The high temperature and pressure kill all microorganisms, including bacterial spores.

e) Coagulase Test (2 marks):

- Definition: The coagulase test is used to differentiate between Staphylococcus aureus (coagulase-
positive) and other staphylococci (coagulase-negative).

- Mechanism: Coagulase enzyme converts fibrinogen to fibrin, causing clot formation. Positive result
indicates *S. aureus*, which can be pathogenic.
f) ELISA (2 marks):

- Definition: Enzyme-linked immunosorbent assay (ELISA) is a biochemical technique used to detect

and quantify antibodies or antigens in a sample.

- Mechanism: The antigen or antibody is captured on a solid surface (e.g., a microplate), followed by
the addition of a substrate that reacts with an enzyme to produce a color change. This color change
is measured to quantify the analyte.

4. Single Most Appropriate Answer (1x5)

i) Food poisoning refers to:

b) Toxin mediated diarrhea

ii) Lymphatic filariasis caused by:

c) Wuchereria bancrofti

iii) What is the best laboratory method to test for HIV infection during the window period?

a. p24 antigen assay

- Explanation: During the window period, which is the early phase of HIV infection when the body
has not yet developed detectable levels of antibodies, the p24 antigen assay is the best method for
detecting HIV. The p24 protein is a part of the HIV virus itself and is detectable in the blood before
antibodies are formed. This test is highly useful in identifying HIV infection during the window
period, typically within 2-3 weeks after exposure.

iv) Which immunoglobulin is normally transported across the placenta?

d. IgG

- Explanation: IgG is the only immunoglobulin that is actively transported across the placenta from
the mother to the fetus. This provides the newborn with passive immunity, protecting it from
infections during the first few months of life. IgM, IgA, and IgE do not cross the placenta.

v) All the viruses belong to Herpesviridae except:

c. Coxsackie virus

- Explanation: The Herpesviridae family includes viruses such as:

- Herpes simplex virus (HSV),

- Epstein-Barr virus (EBV),

- Cytomegalovirus (CMV),

- And others like Varicella-zoster virus (VZV).

Coxsackie virus, however, is a member of the Picornaviridae family, not Herpesviridae.

Coxsackieviruses are associated with conditions like hand, foot, and mouth disease, and viral
myocarditis.

1ST INTERNAL
1. Antigen-Antibody Reactions

#Properties and Types of Antigen-Antibody Reactions:

Antigen-antibody reactions are fundamental to the immune response and are critical in diagnosing
various infections, monitoring immune system activity, and producing therapeutic antibodies. The
main properties of these reactions include:

- Specificity: Antibodies recognize and bind specifically to the antigen that induced their production.

- Affinity: The strength of the binding between an antigen and its corresponding antibody.

- Avidity: The overall strength of the antigen-antibody interaction, which takes into account both the
affinity and the number of binding sites.

- Reversibility: The binding between antigens and antibodies is reversible under certain conditions.

Types of Antigen-Antibody Reactions:

1. Precipitation Reaction: Involves the formation of an insoluble complex when soluble antigens
react with antibodies. This can occur in a test tube, where the antigen-antibody complex forms a
visible precipitate.

2. Agglutination Reaction: Occurs when antibodies cause antigens (usually on the surface of cells) to
clump together. This is a common reaction in blood typing.

3. Complement Fixation: When an antigen-antibody complex binds to the complement system,

leading to the activation of complement proteins and eventually cell lysis.

4. Neutralization: The binding of an antibody to a toxin or virus that prevents its biological activity.
5. Fluorescent Antibody Test: Antibodies tagged with fluorescent markers bind to the corresponding
antigen, allowing for visualization under a fluorescence microscope.

#Principle, Types, and Applications of ELISA:

Principle:

Enzyme-Linked Immunosorbent Assay (ELISA) is based on the principle of antigen-antibody

interaction, where an enzyme-labeled antibody or antigen is used to detect the presence of the
corresponding antigen or antibody. The enzyme produces a color change in the presence of a
substrate, which can be measured spectrophotometrically.

Types of ELISA:

1. Direct ELISA: The antigen is immobilized on the plate, and a labeled antibody is used to detect it.

2. Indirect ELISA: The antigen is immobilized, and an unlabeled primary antibody is added, followed
by a labeled secondary antibody that binds to the primary antibody.

3. Sandwich ELISA: Involves the immobilization of an antibody, followed by the antigen and then the
addition of another labeled antibody that binds to a different epitope on the antigen.

4. Competitive ELISA: Involves competition between a labeled antigen and an unlabeled sample
antigen for binding to a limited amount of antibody.

Applications of ELISA:

- Diagnostic Testing: ELISA is widely used for detecting infectious diseases (e.g., HIV, Hepatitis, Lyme
disease).

- Food and Environmental Testing: Detects allergens, toxins, and contaminants in food or water.

- Hormonal Assays: Used to measure levels of hormones like insulin, human chorionic gonadotropin
(hCG), or thyroid hormones.

- Cancer Markers: Detection of specific proteins or biomarkers associated with cancer.

- Vaccine Development: Used to assess immune responses by measuring antibodies against vaccines.

2. Sterilization and Disinfection

#Definitions:

- Sterilization: The process of eliminating all forms of microbial life, including bacteria, viruses, fungi,
and spores. This is achieved through physical, chemical, or mechanical methods.

- Disinfection: The process of eliminating or reducing harmful microorganisms, but not necessarily all
microbes, especially resistant spores. Disinfection is less stringent than sterilization.

#Methods of Sterilization:
1. Heat Sterilization:

- Autoclaving (Steam under pressure): The most common method, which uses high-pressure steam
to kill microorganisms. It operates at temperatures of 121°C for 15-20 minutes.

- Dry Heat: Involves sterilizing items by exposing them to hot air (160-170°C for 2 hours).

2. Filtration: Removes microorganisms from liquids or gases through a filter with a pore size that
captures bacteria or viruses.

3. Radiation: Gamma rays, X-rays, or UV light can sterilize by damaging the DNA or cellular structures
of microbes.

4. Chemical Sterilization: Involves the use of chemical agents such as ethylene oxide or hydrogen
peroxide to sterilize heat-sensitive materials.

5. Gas Sterilization: Ethylene oxide gas is used to sterilize items that cannot withstand high
temperatures, like medical devices and plastics.

#Methods of Disinfection:

1. Physical Methods:

- Heat: Boiling or pasteurization (at lower temperatures).

- UV Radiation: Kills microorganisms by damaging their DNA.

2. Chemical Methods:

- Alcohol (e.g., ethanol or isopropyl alcohol) for skin disinfection.

- Chlorine compounds (e.g., bleach) for surface disinfection.

- Phenolic compounds for disinfecting hard surfaces.

3. UV Light: Used for disinfecting water or air in hospitals, laboratories, and other settings.

#Autoclave (Brief Description):

An autoclave is a device used for sterilization using pressurized steam at a temperature of 121°C,
typically for 15–20 minutes. It is widely used in medical and laboratory settings for sterilizing surgical
instruments, glassware, and culture media. Autoclaving effectively kills bacteria, viruses, fungi, and
spores by penetrating porous materials and ensuring complete sterilization.
3. Short Notes on (Any Five)

1. Bacterial Growth Curve:

The bacterial growth curve represents the changes in the number of bacterial cells over time. It has
four main phases:

- Lag Phase: Cells are metabolically active but not dividing.

- Log (Exponential) Phase: Cells divide at a constant rate, leading to exponential growth.

- Stationary Phase: Growth rate equals the death rate due to depletion of nutrients and
accumulation of waste products.

- Death (Decline) Phase: Cells die at a faster rate than new cells are formed due to nutrient
exhaustion and toxicity from waste products.

2. Koch's Postulates:

Koch's postulates are a set of criteria used to establish a causal relationship between a microbe and
a disease:

1. The microorganism must be present in all cases of the disease.

2. The microorganism must be isolated from the host and grown in pure culture.

3. The microorganism must cause the disease when introduced into a healthy, susceptible host.

4. The microorganism must be reisolated from the experimentally infected host.

3. Coagulase Test:

The coagulase test is used to identify *Staphylococcus aureus* by detecting the production of
coagulase, an enzyme that causes blood plasma to clot. The test involves adding the bacterial culture
to plasma, and if clotting occurs, the organism is coagulase-positive, indicating it is likely *S. aureus*.

4. Dental Caries:

Dental caries (tooth decay) is caused by the demineralization of tooth enamel due to the acid
produced by bacteria (e.g., *Streptococcus mutans*) that ferment sugars. The acidic environment
leads to the dissolution of mineral salts in the enamel, resulting in cavities. Prevention includes good
oral hygiene, fluoride use, and reducing sugar intake.

5. Conjugation:
Conjugation is a type of horizontal gene transfer in bacteria where genetic material is transferred
from one bacterium to another through direct contact. This process often involves a plasmid (a small
DNA molecule) and plays a crucial role in the spread of antibiotic resistance.

4. Write Briefly on the Following

a. Two Selective Culture Media for *Corynebacterium diphtheriae*:

1. Loeffler's Medium: A selective medium used for the cultivation of *Corynebacterium diphtheriae*;
it promotes the formation of characteristic metachromatic granules.

2. Cystine-Tellurite Agar: Selective for *Corynebacterium diphtheriae*, this medium contains

tellurite which inhibits most bacteria other than *C. diphtheriae*.

b. Two Anaerobic Culture Media:

1. Thioglycollate Agar: Contains reducing agents that create an anaerobic environment, used to
culture anaerobic bacteria.

2. Cooked Meat Medium: A rich medium for the growth of anaerobes, often used in microbiology for
the cultivation of strict anaerobes.

c. Two Biological Effects of Complement:

1. Opsonization: Complement proteins (e.g., C3b) bind to pathogens, enhancing their phagocytosis
by immune cells.

2. Cell Lysis: The membrane attack complex (MAC) formed by complement proteins punctures the
membrane of pathogens, leading to cell lysis.

d. Five Golden Moments of Hand Hygiene:

1. Before patient contact.

2. Before aseptic tasks (e.g., inserting catheters).

3. After body fluid exposure risk.

4. After patient contact.

5. After touching patient surroundings.

e. Two Nonsuppurative Complications of Streptococcus pyogenes:

1. Rheumatic Fever: An inflammatory disease that can affect the heart, joints, and nervous

2ND INTERNAL
1. Hypersensitivity Reactions

#Definition and Classification of Hypersensitivity Reactions:

Hypersensitivity refers to an exaggerated immune response to an antigen, which results in tissue

damage. This immune response can be classified into four types based on the mechanism and the
time course of the reaction:

1. Type I (Immediate Hypersensitivity) – IgE-mediated reaction (e.g., allergies)

2. Type II (Cytotoxic Hypersensitivity) – IgG/IgM-mediated reaction involving cell destruction (e.g.,

hemolytic anemia)

3. Type III (Immune Complex-Mediated Hypersensitivity) – Formation of immune complexes leading

to tissue damage (e.g., systemic lupus erythematosus)

4. Type IV (Delayed-Type Hypersensitivity) – T-cell mediated reaction (e.g., contact dermatitis)

#Type I Hypersensitivity (Immediate Hypersensitivity):

Principle: Type I hypersensitivity is mediated by immunoglobulin E (IgE) antibodies, which bind to

mast cells and basophils. Upon re-exposure to the allergen, the allergen cross-links IgE on these cells,
triggering the release of histamine and other inflammatory mediators (e.g., leukotrienes,
prostaglandins).

Mechanism:

- Sensitization phase: Initial exposure to an allergen results in the activation of helper T cells (Th2
type), which promote IgE production by B cells. The IgE antibodies bind to the surface of mast cells
and basophils.

- Effector phase: On subsequent exposure to the same allergen, it cross-links the IgE on sensitized
mast cells, leading to the release of histamine, cytokines, and other mediators. This results in acute
inflammation and the typical symptoms of an allergic reaction.

Clinical Features:

- Anaphylaxis: A severe, systemic reaction involving widespread vasodilation, bronchoconstriction,

and shock. Symptoms include difficulty breathing, hypotension, and swelling (e.g., angioedema).

- Allergic rhinitis: Sneezing, runny nose, itching, and congestion due to nasal mucosal inflammation.

- Asthma: Inflammation and narrowing of the airways lead to wheezing, coughing, and shortness of
breath.

- Urticaria (hives): Raised, red, itchy welts on the skin due to local histamine release.

- Food allergies: Reactions ranging from mild symptoms (itching, swelling) to severe anaphylaxis.
Laboratory Diagnosis:

- Skin prick test: Small amounts of allergens are introduced into the skin. Positive results lead to
localized wheal and flare reactions.

- Serum IgE levels: Elevated IgE levels are associated with allergic conditions.

- Specific IgE tests: Detect IgE antibodies against particular allergens (e.g., pollen, peanuts).

2. Classification of Mycobacteria and Pathogenesis of Pulmonary Tuberculosis

#Classification of Mycobacteria:

*Mycobacteria* are divided into two main categories based on their clinical and microbiological
properties:

1. Mycobacterium tuberculosis complex:

- Mycobacterium tuberculosis (the primary causative agent of tuberculosis).

- Mycobacterium bovis (rare, causes tuberculosis in cattle but can infect humans).

- Mycobacterium africanum (a variant found primarily in West Africa).

2. Non-tuberculous mycobacteria (NTM) or Atypical Mycobacteria:

- Includes species such as *Mycobacterium avium complex* (MAC), *Mycobacterium kansasii*,

*Mycobacterium marinum*, *Mycobacterium abscessus*, etc. These are usually opportunistic
pathogens.

#Pathogenesis of Pulmonary Tuberculosis (TB):

Pulmonary tuberculosis is primarily caused by *Mycobacterium tuberculosis* and involves the

following stages:

1. Inhalation of Mycobacteria: TB is transmitted via aerosol droplets from an infected person. The
bacteria reach the alveoli in the lungs.

2. Phagocytosis by Alveolar Macrophages: The bacteria are engulfed by macrophages but are not
killed due to the mycolic acid in their cell walls, which protects them from lysosomal enzymes.

3. Formation of Granulomas: The body’s immune response attempts to contain the bacteria by
forming granulomas (a collection of macrophages, lymphocytes, and fibroblasts). In the center of the
granuloma, a caseous necrosis (cheese-like appearance) may develop.

4. Latent TB: In most healthy individuals, the immune system can contain the infection, and the
bacteria remain dormant in the granulomas.

5. Active TB Disease: If the immune system weakens (e.g., in immunocompromised individuals), the
bacteria can reactivate and cause active disease, leading to symptoms such as chronic cough, fever,
weight loss, night sweats, and hemoptysis (coughing up blood).
#Laboratory Diagnosis of Pulmonary Tuberculosis:

1. Microscopy: Acid-fast bacilli (AFB) can be observed in sputum samples using the Ziehl-Neelsen
stain or the fluorescence staining method.

2. Culture: The gold standard for diagnosing TB is the culture of *M. tuberculosis* from sputum or
other clinical specimens using Lowenstein-Jensen (LJ) medium or automated liquid culture systems.

3. Polymerase Chain Reaction (PCR): PCR tests, such as Xpert MTB/RIF, can detect *M. tuberculosis*
DNA and assess for rifampin resistance.

4. Chest X-ray: Radiological evidence of lung damage, such as cavitations or infiltrates, is suggestive
of active pulmonary tuberculosis.

5. Tuberculin Skin Test (TST) or Mantoux Test: Measures the delayed hypersensitivity response to
purified protein derivative (PPD) from *M. tuberculosis*. It is used for screening but not for
diagnosing active TB.

3. Short Notes on (Any Five)

a. Difference between Primary and Secondary Immune Response:

- Primary Immune Response:

- Occurs when the immune system is exposed to an antigen for the first time.

- Characterized by a lag period of several days (5–10 days) before detectable antibodies appear.

- Initially, IgM antibodies are produced, followed by IgG.

- The response is slower and less intense.

- Secondary Immune Response:

- Occurs upon subsequent exposure to the same antigen.

- The immune system reacts more rapidly and more strongly due to the presence of memory B and
T cells.

- Predominantly IgG antibodies are produced, and the response is faster and more efficient.

b. Oral Candidiasis:

Oral candidiasis, also known as thrush, is a fungal infection caused by *Candida* species, particularly
*Candida albicans*. It manifests as white, creamy lesions on the tongue, gums, palate, and inner
cheeks. Risk factors include immunosuppression (e.g., HIV/AIDS), diabetes, antibiotic use, dentures,
and poor oral hygiene. Treatment typically involves antifungal medications such as nystatin or
fluconazole.
c. Streptococcal Pharyngitis:

Streptococcal pharyngitis, or strep throat, is an infection of the throat and tonsils caused by
*Streptococcus pyogenes* (group A Streptococcus). Symptoms include sore throat, fever, headache,
and swollen lymph nodes. Diagnosis is confirmed by throat culture or rapid antigen test. Treatment
with antibiotics, usually penicillin or amoxicillin, reduces the risk of complications like rheumatic
fever.

d. Risk Factors for Developing Opportunistic Fungal Infections:

- Immunocompromised states (e.g., HIV/AIDS, cancer, organ transplantation).

- Antibiotic use (which disrupts normal microbiota and allows fungal overgrowth).

- Diabetes mellitus (especially when poorly controlled).

- Corticosteroid use (which suppresses immune function).

- Indwelling medical devices (e.g., catheters, prosthetic devices) that increase the risk of infection.

e. Toxins Produced by Staphylococcus aureus:

- Alpha toxin: Forms pores in the cell membranes of host cells, leading to cell lysis and tissue
damage.

- Beta toxin: Hydrolyzes sphingomyelin in cell membranes, causing damage to red blood cells and
tissues.

- Exfoliative toxins (ETA and ETB): Responsible for the skin sloughing in conditions like
*Staphylococcal scalded skin syndrome.

- Toxic shock syndrome toxin (TSST-1): A superantigen that stimulates massive cytokine release,
leading to shock and multi-organ failure.

- Enterotoxins: Cause food poisoning by inducing vomiting and diarrhea.

4. Write Briefly on the Following

a. Diseases Produced by S. aureus:

1. Skin Infections: Boils, impetigo, cellulitis, and abscesses.

2. Food Poisoning: Caused by ingestion of enterotoxin-producing strains.

3. Pneumonia: Particularly in individuals with underlying lung disease.

4. Toxic Shock Syndrome: A severe systemic condition caused by TSST-1.

5. Endocarditis:

Infection of the heart valves.

6. Osteomyelitis: Bone infection, often following trauma or surgery.

b. Two Beta-Hemolytic Streptococcus:

1. Streptococcus pyogenes (Group A): Causes pharyngitis, scarlet fever, rheumatic fever, and
necrotizing fasciitis.

2. Streptococcus agalactiae (Group B): Common cause of neonatal infections, including sepsis and
meningitis.

c. Two Major or Professional Antigen-Presenting Cells (APCs):

1. Dendritic cells: Highly efficient in antigen uptake and presentation to naïve T cells.

2. Macrophages: Present antigens to T cells after phagocytosing pathogens.

d. Two Effector Functions of Antibody-Mediated Immunity (AMI):

1. Neutralization: Antibodies bind to pathogens or toxins, preventing them from interacting with
host cells.

2. Opsonization: Antibodies enhance the phagocytosis of pathogens by immune cells like

macrophages and neutrophils.

e. Two Non-Albicans Species of Candida:

1. Candida glabrata: Known for its resistance to antifungal treatment and common in
immunocompromised patients.

2. Candida krusei: Often resistant to fluconazole and associated with bloodstream infections in
immunocompromised individuals.

3RD INTERNAL
1. Structure, Pathogenesis, and Laboratory Diagnosis of HIV

#Structure of HIV:

HIV (Human Immunodeficiency Virus) is a retrovirus with the following key features:

- **Envelope**: The virus is enveloped with a lipid bilayer derived from the host cell membrane.
Embedded in this envelope are viral glycoproteins, primarily **gp120** and **gp41**, which are
essential for binding to host cell receptors.

- **Capsid**: The core of the virus contains a protein shell made of the **p24** protein, which is
critical for the virus's structural integrity.
- **RNA Genome**: HIV contains two single-stranded RNA molecules as its genetic material, which
is reverse-transcribed into DNA once inside the host cell.

- **Reverse Transcriptase**: HIV carries the enzyme reverse transcriptase, which converts its RNA
genome into DNA after entering the host cell.

- **Integrase**: This enzyme is involved in integrating the viral DNA into the host cell genome.

#Pathogenesis of HIV:

1. **Entry into Host Cells**: HIV primarily infects CD4+ T lymphocytes, macrophages, and dendritic
cells. The virus binds to the **CD4 receptor** on the host cell surface, and the interaction with the
**CCR5** or **CXCR4** co-receptors facilitates fusion with the cell membrane.

2. **Reverse Transcription**: After fusion, the viral RNA is reverse-transcribed into DNA by the viral
reverse transcriptase enzyme.

3. **Integration into Host Genome**: The newly synthesized viral DNA is transported into the
nucleus, where **integrase** incorporates it into the host's DNA, forming a provirus.

4. **Replication**: The host cell machinery transcribes and translates the viral genome, producing
viral RNA and proteins, which are assembled into new virions.

5. **Budding and Spread**: Newly formed HIV particles bud off from the host cell, which may cause
cell death. The virus spreads to other immune cells, leading to systemic immune dysfunction.

6. **Immune System Dysfunction**: Over time, the progressive destruction of CD4+ T cells leads to
immunodeficiency. This weakens the immune system, making the body susceptible to opportunistic
infections and certain cancers.

#Laboratory Diagnosis of HIV:

1. **Serological Tests**:

- **Enzyme-Linked Immunosorbent Assay (ELISA)**: Used to detect antibodies against HIV. It is

highly sensitive and used for initial screening.

- **Western Blot**: Used to confirm a positive ELISA result. It detects specific HIV proteins (e.g.,
gp160, gp120, and p24).

- **Rapid HIV Tests**: Immunoassays that provide quick results by detecting HIV antibodies or
antigens.
2. **Nucleic Acid Tests (NAT)**: These detect HIV RNA or DNA in the blood. They are used to
diagnose early infection (before antibody development) or to monitor viral load during treatment.

3. **CD4+ T Cell Count**: This test measures the number of CD4+ T lymphocytes in the blood. A low
CD4 count is indicative of advanced HIV infection and progression to AIDS.

4. **HIV Viral Load**: Quantification of HIV RNA in the blood is an important marker of active
infection and is used to monitor the effectiveness of antiretroviral therapy (ART).

5. **PCR (Polymerase Chain Reaction)**: Used to detect HIV RNA or proviral DNA, especially in
infants (to distinguish between maternal antibodies and the infant's infection).

2. **Causative Agents of Malaria, Laboratory Diagnosis, and Complications of Plasmodium

falciparum Malaria**

#Causative Agents of Malaria:

Malaria is caused by protozoan parasites of the genus *Plasmodium*. The four main species
responsible for malaria in humans are:

1. **Plasmodium falciparum**: The most virulent and common species causing severe malaria.

2. **Plasmodium vivax**: Causes a milder form of malaria, but can relapse due to dormant liver
stages (hypnozoites).

3. **Plasmodium ovale**: Similar to *P. vivax*, with the potential for relapse.

4. **Plasmodium malariae**: Causes a milder form of malaria with a longer periodicity in fever
cycles.

#Laboratory Diagnosis of Malaria:

1. **Microscopic Examination**:

- **Thick Blood Smear**: Used to detect the presence of *Plasmodium* parasites. The thick smear
concentrates the blood, making it easier to detect parasites.

- **Thin Blood Smear**: Used for species identification by examining the morphology of the
parasite.

2. **Rapid Diagnostic Tests (RDTs)**: These detect specific *Plasmodium* antigens in the blood and
provide rapid results, useful in settings with limited laboratory infrastructure.
3. **Polymerase Chain Reaction (PCR)**: A highly sensitive test that can detect low levels of
parasites and identify the *Plasmodium* species. It is especially useful for cases with mixed
infections or low parasitemia.

4. **Serological Tests**: Detect antibodies or antigens, but these are not typically used for diagnosis
of active malaria.

#Complications of *Plasmodium falciparum* Malaria:

*Plasmodium falciparum* is the most dangerous species and can cause severe complications,
including:

1. **Cerebral Malaria**: Involves sequestration of parasitized red blood cells in the small blood
vessels of the brain, leading to encephalopathy, seizures, and coma.

2. **Severe Anemia**: Caused by the destruction of red blood cells by the parasite, leading to
significant blood loss.

3. **Acute Renal Failure**: Due to the destruction of red blood cells, leading to hemoglobinuria and
kidney damage.

4. **Respiratory Distress**: Can result from metabolic acidosis, pulmonary edema, or acute
respiratory distress syndrome (ARDS).

5. **Hypoglycemia**: *Plasmodium falciparum* infection can cause a significant drop in blood

glucose levels, especially in pregnant women or children.

6. **Multisystem Organ Failure**: In severe cases, the parasite can cause dysfunction in multiple
organs, including the liver, kidneys, and heart.

3. **Short Notes on (Any Five)**

a. **Autoimmunity**:

Autoimmunity occurs when the body's immune system mistakenly attacks its own tissues as if they
were foreign. It is often triggered by genetic, environmental, or hormonal factors. Examples of
autoimmune diseases include rheumatoid arthritis, systemic lupus erythematosus (SLE), and
multiple sclerosis.

b. **VDRL (Venereal Disease Research Laboratory Test)**:

The VDRL test is a non-treponemal serological test used for the detection of **syphilis**. It detects
antibodies against cardiolipin, a lipid released from damaged cells, rather than the causative
bacterium *Treponema pallidum*. It is a screening test and is often followed by a more specific test,
such as the **FTA-ABS** test, to confirm the diagnosis.
c. **Laboratory Diagnosis of Leprosy**:

- **Skin Smear**: A sample from a skin lesion is examined under a microscope after staining with
**Ziehl-Neelsen stain**. The presence of **acid-fast bacilli (AFB)** confirms the diagnosis.

- **Skin Biopsy**: In cases where a smear is negative, a biopsy can help in diagnosing leprosy by
detecting granulomas and the presence of *Mycobacterium leprae*.

- **PCR**: Molecular techniques can detect *M. leprae* DNA in tissue or blood samples.

d. **Difference between Actinomycosis and Nocardiosis**:

- **Actinomycosis**:

- Caused by *Actinomyces* species (Gram-positive anaerobic bacteria).

- Characterized by **sulfur granules** in pus, and often results in abscess formation in the face,
jaw, and chest.

- Often associated with chronic infection following trauma or poor dental hygiene.

- **Nocardiosis**:

- Caused by *Nocardia* species (Gram-positive, partially acid-fast bacteria).

- Affects the lungs, skin, and brain, and can present with **granulomatous abscesses**.

- It is more common in immunocompromised individuals and presents with a more acute,

disseminated infection.

e. **Graft-versus-Host Reaction (GVHR)**:

GVHR occurs after hematopoietic stem cell or organ transplantation, where the transplanted
**immune cells** recognize the recipient’s tissues as foreign and mount an immune response
against them. This leads to damage of the recipient's organs, particularly the skin, liver, and
gastrointestinal tract. GVHR can range from mild to severe and can be life-threatening.

4. **Write Briefly on the Following**

a. **Two Viruses That Have Previously Caused Pandemics**:

1. **Influenza Virus (H1N1)**: The 2009 H1N1 "swine flu" pandemic caused global illness and
mortality.

2. **SARS-CoV-2 (COVID-19)**: The novel coronavirus responsible for the ongoing global pandemic
that began in 2019.
b. **Two Methods of Virus Isolation**:

1. **Cell Culture**: Viruses are grown in suitable mammalian or insect cell lines, and cytopathic
effects (CPE) are observed to confirm the presence of the virus.

2. **Embryonated Egg Inoculation**: Some viruses, such as influenza and yellow fever, are cultured
in fertilized chicken eggs, where they replicate.

c. **Two Clinical Variants of Leprosy**:

1. **Tuberculoid Leprosy**: Characterized by a few well-defined skin lesions, with granulomatous

inflammation and strong immune response.

2. **Lepromatous Leprosy**: A more severe form, with widespread skin lesions, nerve damage, and
no effective immune response.

d. **Two Immunodeficiency Diseases**:

1. **Severe Combined Immunodeficiency (SCID)**: A genetic disorder resulting in severe defects in

both T-cell and B-cell immunity.

2. **Common Variable Immunodeficiency (CVID)**: A disorder of antibody production leading to

recurrent infections.

e. **Two Spore-Bearing Anaerobes**:

1. **Clostridium tetani**: Causes tetanus, characterized by muscle spasms.

2. **Clostridium botulinum**: Produces botulinum toxin, which causes botulism, a form of food
poisoning.