Molecular Biology Techniques in Microbiology Practice Questions

Q: Two different specimens from two patients are received. Which of the following techniques can confirm they are from different individuals (in the context of Mycobacterium tuberculosis)?

RFLP. ***Correct: RFLP*** - **RFLP** (Restriction Fragment Length Polymorphism), particularly using the **IS6110 insertion sequence**, is the traditional gold standard tool for **epidemiological strain typing** of *M. tuberculosis*. - Confirmation that two specimens are from different individuals relies on visualizing two distinct banding patterns, indicating different arrangements of the **IS6110** elements in their respective genomes. - RFLP has **high discriminatory power** for differentiating between strains and establishing epidemiological links. *Incorrect: Mutation analysis* - This technique primarily focuses on detecting **point mutations** in specific genes (e.g., *rpoB*, *katG*) to determine drug susceptibility or resistance patterns. - While differences in resistance profiles suggest different strains, it lacks the **high discriminatory power** of RFLP or VNTR for broad epidemiological linkage assessment. *Incorrect: Pyrosequencing* - Pyrosequencing is a rapid method of DNA sequencing often used to detect **single nucleotide polymorphisms (SNPs)** or specific mutations, such as those related to drug resistance. - It is utilized for sequencing short regions and is not the primary technique for comparing the **large-scale genomic structures** necessary for definitive strain fingerprinting. *Incorrect: RAPD* - **RAPD** (Random Amplification of Polymorphic DNA) is an older PCR-based method that uses arbitrary primers to generate a pattern of amplified DNA fragments. - While used for strain comparison in some bacteria, RAPD is generally considered **less reproducible** and has **lower discriminatory power** than RFLP or VNTR for *M. tuberculosis* typing.

Question 1

A microbiologist is working with PCR and wishes to quantify the PCR by monitoring the amplification process while the PCR is ongoing. Which of the following types of PCR should be used?

Accepted Answer

Real-Time PCR (qPCR)

Answer

Standard PCR

Answer

Reverse Transcriptase PCR

Answer

Multiplex PCR

Question 2

Two different specimens from two patients are received. Which of the following techniques can confirm they are from different individuals (in the context of Mycobacterium tuberculosis)?

Accepted Answer

RFLP

Answer

Pyrosequencing

Answer

Mutation analysis

Answer

RAPD

Molecular Biology Techniques in Microbiology — MCQs

Molecular Biology Techniques in Microbiology — MCQs

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