Laboratory Diagnosis of Parasitic Infections — MCQs
In a patient presented with a fever and a positive filarial antigen test, what is the next appropriate method of management?
A frequent traveler presented with 4 days of continuous fever, abdominal pain, and bradycardia. What is the best diagnostic test to confirm the pathogen?
What is the best investigation for identifying malaria species?
Which PCR technique is best suited for identifying a syndrome with multiple causative agents?
JSB stain is used for which parasite?
What is the most sensitive diagnostic method for detecting Trichomonas vaginalis?
A Giemsa stain of a thin peripheral blood smear is prepared. Which of the following cannot be diagnosed?
Which of the following is a xenodiagnostic method?
Hanging drop method is used for:
Statement 1 - A 59-year-old patient presents with flaccid bullae. Histopathology shows a suprabasal acantholytic split. Statement 2 - The row of tombstones appearance is diagnostic of Pemphigus vulgaris.
Laboratory Diagnosis of Parasitic Infections Indian Medical PG Practice Questions and MCQs
Question 1: In a patient presented with a fever and a positive filarial antigen test, what is the next appropriate method of management?
- A. Bone marrow biopsy
- B. DEC provocation test
- C. Detection of microfilariae in the blood smear (Correct Answer)
- D. Ultrasound of the scrotum
Explanation: ***Detection of microfilariae in the blood smear*** - A positive **filarial antigen test** indicates the presence of adult worms, and the next step is to confirm active infection by identifying **microfilariae**. [1] - **Nocturnal blood samples** are crucial because microfilariae of *Wuchereria bancrofti* and *Brugia malayi* exhibit **nocturnal periodicity**, meaning they are most abundant in peripheral blood between 10 PM and 2 AM. [1] *Bone marrow biopsy* - This procedure is typically used to diagnose **hematological disorders**, such as leukemia or lymphoma, or investigate causes of unexplained fever, but it is not indicated for filariasis. - While filariasis can rarely lead to **eosinophilia**, a bone marrow biopsy is not a diagnostic tool for filarial infection itself. *DEC provocation test* - The **diethylcarbamazine (DEC) provocation test** is used to bring out microfilariae into the peripheral blood during the daytime for species that exhibit nocturnal periodicity. [1] - However, it carries a risk of severe adverse reactions due to rapid killing of microfilariae, especially in cases of heavy infection, and is generally avoided when antigen tests are positive. [1] *Ultrasound of the scrotum* - Scrotal ultrasound can detect the characteristic "filarial dance sign" (motile adult worms) in the **lymphatic vessels of the scrotum and epididymis**, confirming lymphatic filariasis. [2] - While useful for assessing advanced disease manifestations like **hydrocele**, it does not quantify microfilaremia or replace the need for microscopic confirmation of circulating microfilariae to guide treatment.
Question 2: A frequent traveler presented with 4 days of continuous fever, abdominal pain, and bradycardia. What is the best diagnostic test to confirm the pathogen?
- A. Widal test
- B. Blood culture (Correct Answer)
- C. Urine culture
- D. Stool culture
Explanation: ***Blood culture*** - **Blood culture** is the most sensitive and specific test for confirming **typhoid fever** in the first week of illness. - The presence of **continuous fever** (step-ladder pattern), **abdominal pain**, and **relative bradycardia** in a traveler strongly suggests typhoid fever caused by *Salmonella Typhi*. *Widal test* - The **Widal test** detects antibodies against *Salmonella Typhi* antigens and is often positive later in the disease course. - It has **limited sensitivity and specificity**, especially in endemic areas or with prior vaccination, leading to false positives and negatives. *Urine culture* - **Urine culture** has a low yield for *Salmonella Typhi*, as bacteria are intermittently shed in urine, usually later in the disease. - It's primarily useful for diagnosing **urinary tract infections** or in chronic carriers of typhoid. *Stool culture* - **Stool culture** yield is higher in the later stages of typhoid fever, as *Salmonella Typhi* is shed in feces. - Its sensitivity is lower than blood culture in the early acute phase when bacteremia is most prominent.
Question 3: What is the best investigation for identifying malaria species?
- A. Thick smear
- B. Thin smear with Giemsa (Correct Answer)
- C. QBC
- D. Thin smear with acridine orange
Explanation: ***Thin smear with Giemsa*** - A **thin smear** allows for the visualization of **parasite morphology** within red blood cells, which is crucial for distinguishing between species of *Plasmodium*. - **Giemsa stain** provides optimal contrast for identifying characteristic features such as **merozoites**, **trophozoites**, **schizonts**, and **gametocytes** of different malaria species. *Thick smear* - A **thick smear** is primarily used for **detecting the presence of malaria parasites** and for quantifying parasite density due to its higher sensitivity. - However, because red blood cells are lysed, it **does not preserve parasite morphology** well, making species identification difficult. *QBC* - **Quantitative Buffy Coat (QBC) analysis** is a rapid method for detecting malaria parasites based on their fluorescence under UV light. - While sensitive for detection, it generally **does not allow for precise species identification** due to the lack of clear morphological detail. *Thin smear with acridine orange* - A **thin smear stained with acridine orange** is used for rapid detection of parasites by fluorescence microscopy. - Similar to QBC, it is **less effective for detailed morphological examination** and specific species identification compared to Giemsa-stained thin smears.
Question 4: Which PCR technique is best suited for identifying a syndrome with multiple causative agents?
- A. RT-PCR
- B. Multiplex PCR (Correct Answer)
- C. Nested PCR
- D. Conventional PCR
Explanation: ***Multiplex PCR*** - **Multiplex PCR** allows for the simultaneous amplification of **multiple DNA targets** in a single reaction, making it ideal for identifying syndromes with numerous potential causative agents. - This method uses **multiple primer pairs** in one reaction tube, each designed to amplify a specific target sequence, thus efficiently detecting various pathogens or genetic markers. *RT-PCR* - **Reverse Transcription PCR (RT-PCR)** is used to detect **RNA targets** by first converting RNA into cDNA, which is then amplified. - While useful for RNA viruses or gene expression studies, it is not primarily designed for simultaneous detection of multiple diverse causative agents in the same way as multiplex PCR. *Nested PCR* - **Nested PCR** uses two sets of primers in sequential reactions to **increase sensitivity and specificity** by reducing non-specific binding. - This technique is generally employed to detect very low copies of a specific target or to overcome issues with non-specific amplification, rather than for identifying multiple different agents concurrently. *Conventional PCR* - **Conventional PCR** amplifies a **single specific DNA target** using one pair of primers per reaction. [1] - It requires separate reactions for each potential causative agent, making it inefficient and labor-intensive when testing for a syndrome with multiple etiologies. **References:** [1] Cross SS. Underwood's Pathology: A Clinical Approach. 6th ed. (Basic Pathology) introduces the student to key general principles of pathology, both as a medical science and as a clinical activity with a vital role in patient care. Part 2 (Disease Mechanisms) provides fundamental knowledge about the cellular and molecular processes involved in diseases, providing the rationale for their treatment. Part 3 (Systematic Pathology) deals in detail with specific diseases, with emphasis on the clinically important aspects., pp. 56-57.
Question 5: JSB stain is used for which parasite?
- A. Kala azar
- B. Sleeping sickness
- C. Malaria
- D. Filaria (Correct Answer)
Explanation: ***Filaria*** - The **JSB stain (Jaswant Singh Battacharya stain)** is a rapid Romanowsky-type stain specifically developed for the diagnosis of **microfilariae** in blood films. - It allows for clear visualization of the sheaths and nuclei of microfilariae, which is crucial for species identification and diagnosis of **filariasis**. *Malaria* - **Giemsa stain** is the gold standard for identifying malaria parasites in thick and thin blood smears, not JSB stain. - Giemsa allows for detailed morphological differentiation of malaria species and stages within **red blood cells**. *Kala azar* - **Kala-azar (visceral leishmaniasis)** is diagnosed by detecting **Leishman bodies (amastigotes)** in bone marrow, splenic, or lymph node aspirates. - Stains like **Giemsa** or **Leishman stain** are traditionally used for visualizing these amastigotes. *Sleeping sickness* - **Sleeping sickness (African trypanosomiasis)** is diagnosed by identifying **trypomastigotes** in blood smears, lymph node aspirates, or cerebrospinal fluid. - **Giemsa stain** is commonly used for the microscopic examination of these specimens to detect the parasites.
Question 6: What is the most sensitive diagnostic method for detecting Trichomonas vaginalis?
- A. Pap smear
- B. Culture
- C. Nucleic acid amplification test (NAAT) (Correct Answer)
- D. Wet mount microscopy
Explanation: ***Nucleic acid amplification test (NAAT)*** - **NAATs** detect **_Trichomonas vaginalis_** DNA or RNA, offering the **highest sensitivity and specificity** among available diagnostic methods. - This method is particularly useful for detecting low parasitic loads and in asymptomatic patients, improving diagnostic accuracy. *Pap smear* - While a **Pap smear** can sometimes incidentally detect **_Trichomonas vaginalis_**, it is not a dedicated or **sensitive diagnostic tool** for this infection. - Its primary purpose is cervical cancer screening, and its sensitivity for trichomoniasis is low, often leading to false negatives. *Culture* - **Culture** was previously considered the **gold standard** but is less sensitive and takes longer (up to 7 days) to yield results compared to NAATs. - Its sensitivity is significantly reduced when parasite loads are low or if samples are not processed promptly. *Wet mount microscopy* - **Wet mount microscopy** allows for the visualization of **motile trichomonads**, but its sensitivity is highly dependent on operator experience and parasitic load. - It has a **sensitivity of 50-70%**, meaning a significant number of infections can be missed.
Question 7: A Giemsa stain of a thin peripheral blood smear is prepared. Which of the following cannot be diagnosed?
- A. Coxiella burnettii (Correct Answer)
- B. Bartonella henselae
- C. Ehrlichia chaffeensis
- D. Toxoplasma gondii
Explanation: ***Coxiella burnettii*** - *Coxiella burnettii* causes **Q fever** and is an **obligate intracellular bacterium** that resides primarily in **tissue macrophages** (lungs, liver, bone marrow), not in circulating blood cells. - It is **not found in peripheral blood smears** because it does not infect circulating leukocytes in significant numbers that would allow microscopic visualization. - Diagnosis requires **serology** (most common), **PCR**, or specialized culture in BSL-3 facilities—direct microscopic visualization in blood smears is not possible. *Bartonella henselae* - Causes **Cat scratch disease** and can invade **red blood cells**, making it potentially visible on Giemsa-stained blood smears, particularly in immunocompromised patients with bacillary angiomatosis or bacteremia. - While difficult and not the primary diagnostic method, it *can* be visualized in peripheral blood, unlike *Coxiella*. *Ehrlichia chaffeensis* - Causes **human monocytotropic ehrlichiosis (HME)** and forms characteristic **morulae** (berry-like clusters) within the cytoplasm of **monocytes**. - These morulae are readily visible on **Giemsa-stained peripheral blood smears** and are a key diagnostic finding, making this condition easily diagnosed by this method. *Toxoplasma gondii* - An **intracellular parasite** whose **tachyzoites** can occasionally be found in **peripheral blood leukocytes** during acute infection, especially in immunocompromised patients. - While rare and not the primary diagnostic method (serology/PCR preferred), tachyzoites *can* be observed in blood smears during active parasitemia.
Question 8: Which of the following is a xenodiagnostic method?
- A. Direct microscopic examination of blood smear for malaria parasites
- B. Injecting a hamster with splenic biopsy for diagnosis of leishmaniasis
- C. Injecting Aedes thorax with blood of a suspected dengue patient (Correct Answer)
- D. Rabbit ileal loop for enterotoxigenic Escherichia coli
Explanation: **Injecting *Aedes* thorax with blood of a suspected dengue patient** - **Xenodiagnosis** involves using a live arthropod vector (or other live animal) to feed on a patient suspected of having a parasitic infection. The subsequent examination of the vector for the pathogen confirms the diagnosis. - In this method, the *Aedes* mosquito, a natural vector for dengue, is used to detect the dengue virus in a patient's blood, fitting the definition of xenodiagnosis. *Direct microscopic examination of blood smear for malaria parasites* - This is a **direct diagnostic method** where the pathogen (malaria parasites) is visualized directly under a microscope from the patient's sample. - It does not involve an intermediate live host or vector, hence it is not xenodiagnosis. *Injecting a hamster with splenic biopsy for diagnosis of leishmaniasis* - This is an example of **animal inoculation** or **culture in vivo**, where an animal is used as a living culture medium to grow the pathogen. - While it uses a live animal, it typically aims to multiply the pathogen for later detection or isolation, rather than using an arthropod vector to 'diagnose' by infection, which is characteristic of xenodiagnosis. *Rabbit ileal loop for enterotoxigenic *Escherichia coli*** - This is a **bioassay** used to detect enterotoxins produced by bacteria, often for research or verification purposes. - It assesses the effect of bacterial toxins on the intestinal mucosa of a live animal, not the detection of a pathogen by allowing an arthropod vector to feed on an infected host.
Question 9: Hanging drop method is used for:
- A. Toxoplasma
- B. Cryptosporidium
- C. Trichomonas (Correct Answer)
- D. Plasmodium
Explanation: ***Trichomonas*** - The **hanging drop method** is a highly effective technique for visualizing the characteristic **motility** of *Trichomonas vaginalis*. - This method allows for the observation of living, unstained organisms directly from clinical samples, making it valuable for rapid diagnosis. *Toxoplasma* - **Toxoplasma gondii** is an intracellular parasite best identified through serological tests for **antibodies** or molecular diagnostics like **PCR**. - It does not exhibit characteristic motility in a hanging drop preparation that would aid in its direct identification. *Cryptosporidium* - **Cryptosporidium** species are typically identified by detecting **oocysts** in stool samples, often using **acid-fast staining** or **immunofluorescence assays**. - Their small size and lack of distinctive motility under a hanging drop method make this technique unsuitable for their diagnosis. *Plasmodium* - **Plasmodium** species, the causative agents of malaria, are diagnosed by visualizing **parasites within red blood cells** on **Giemsa-stained blood smears**. - The hanging drop method would not effectively identify these intracellular parasites for malaria diagnosis.
Question 10: Statement 1 - A 59-year-old patient presents with flaccid bullae. Histopathology shows a suprabasal acantholytic split. Statement 2 - The row of tombstones appearance is diagnostic of Pemphigus vulgaris.
- A. Statements 1 & 2 are correct, 2 is not explaining 1 (Correct Answer)
- B. Statements 1 and 2 are correct and 2 is the correct explanation for 1
- C. Statements 1 and 2 are incorrect
- D. Statement 1 is incorrect
Explanation: ***Correct: Statements 1 & 2 are correct, 2 is not explaining 1*** **Analysis of Statement 1:** - A 59-year-old patient with **flaccid bullae** and **suprabasal acantholytic split** on histopathology is the classic presentation of **Pemphigus vulgaris** - The flaccid (easily ruptured) nature of bullae distinguishes it from tense bullae seen in bullous pemphigoid - The suprabasal location of the split (just above the basal layer) with acantholysis (loss of cell-to-cell adhesion) is pathognomonic - **Statement 1 is CORRECT** ✓ **Analysis of Statement 2:** - The **"row of tombstones" or "tombstone appearance"** is indeed a diagnostic histopathological feature of Pemphigus vulgaris - This appearance results from basal keratinocytes remaining attached to the basement membrane while suprabasal cells separate due to acantholysis - The intact basal cells standing upright resemble a row of tombstones - **Statement 2 is CORRECT** ✓ **Does Statement 2 explain Statement 1?** - Statement 2 describes a **histopathological appearance** (tombstone pattern) that is a **consequence** of the suprabasal split - However, it does NOT explain the **underlying cause** of the flaccid bullae or the suprabasal split - The true explanation involves **IgG autoantibodies against desmoglein 3 (and desmoglein 1)**, which attack intercellular adhesion structures (desmosomes), causing **acantholysis** - Therefore, **Statement 2 does NOT explain Statement 1** ✗ *Incorrect: Statement 2 is the correct explanation for Statement 1* - While both statements describe features of Pemphigus vulgaris, the tombstone appearance is a descriptive finding, not an explanatory mechanism *Incorrect: Statements 1 and 2 are incorrect* - Both statements are medically accurate descriptions of Pemphigus vulgaris features *Incorrect: Statement 1 is incorrect* - Statement 1 correctly describes the cardinal clinical and histopathological features of Pemphigus vulgaris
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