Deck 14: Molecular Techniques

Question 1

The PCR technique can be modified to study ribonucleic acid (RNA)as well as deoxyribonucleic acid (DNA).

Answer 1

True

Question 2

DNA can be amplified by:

A) in vitro amplification.
B) cloning.
C) synthesis of proteins.
D) both a and b

Answer 2

both a and b

Question 3

Clinical outcomes of PCR can lead to:

A) detection of gene mutations that signify the early development of cancer.
B) identification of viral DNA associated with specific cancers.
C) detection of genetic mutations associated with various diseases.
D) all of the above.

Answer 3

all of the above.

Question 4

Multiplex PCR is limited by the number of primers that can be included in a single reaction.

Question 5

MATCHING
Match the technique with an appropriate description (use each answer only once).

Nucleic acid sequence amplification

A)This amplifies target nucleic acid without the use of a thermocycler.A double-stranded DNA fragment is created and becomes the target for exponential amplification.
B)An isothermal assay that targets either DNA or RNA but generates RNA as its amplified product.
C)Oligonucleotide pairs hybridize to target sequences within the gene or the cryptic plasmid.
D)Only RNA is targeted for amplification.

Question 6

The difference between transcription-mediated amplification (TMA)and nucleic acid sequence-based amplification (NASBA)is that:

A) only RNA is targeted for amplification in NASBA.
B) NASBA is not useful for HIV.
C) only DNA is tangled for amplification in NASBA.
D) NASBA is the method of choice for the detection of microorganisms such as Mycobacterium tuberculosis.

Question 7

Amplification of nucleic acid fragments from a different target is performed with:

A) PCR.
B) reverse transcriptase-PCR (RT-PCR).
C) multiplex PCR.
D) real-time PCR.

Question 8

Real-time PCR:

A) is less susceptible to amplicon contamination.
B) is less accurate in quantifying the initial copy number.
C) does not require a thermocycler.
D) quantifies specific RNA sequences.

Question 9

The advantages of molecular testing include all the following except:

A) faster turnaround time.
B) decreased sensitivity.
C) smaller specimen volume required.
D) increased specificity.

Question 10

The gold standard of molecular testing is:

A) DNA sequencing.
B) FISH.
C) reverse transcriptase.
D) nephelometry.

Question 11

The clinical application of the analysis of amplification products includes:

A) genetic testing.
B) hematopathology diagnosis and monitoring.
C) identification of infectious agents.
D) all of the above.

Question 12

MATCHING
Match the technique with an appropriate description (use each answer only once).

Transcription-mediated amplification

A)This amplifies target nucleic acid without the use of a thermocycler.A double-stranded DNA fragment is created and becomes the target for exponential amplification.
B)An isothermal assay that targets either DNA or RNA but generates RNA as its amplified product.
C)Oligonucleotide pairs hybridize to target sequences within the gene or the cryptic plasmid.
D)Only RNA is targeted for amplification.

Question 13

All the following are true in regard to the polymerase chain reaction (PCR)except:

A) it amplifies low levels of specific DNA sequences.
B) the target sequence to be amplified can be unknown.
C) two short DNA primers are used.
D) oligonucleotides act as a template.

Question 14

Reverse-transcriptase polymerase chain reaction (RT-PCR)applications include identification of:

A) Treponema pallidum (spirochete bacteria).
B) HIV.
C) HCV.
D) both b and c

Question 15

Arrange the following steps in the enzymatic process of the PCR cycle in the correct chronologic order:
1)Primer annealing
2)DNA denaturation
3)Extension of the primed DNA sequence

A) 1,2,3
B) 2,1,3
C) 2,3,1
D) 3,2,1

Question 16

Multiplex PCR is limited by specific nucleic acid amplification.

Question 17

The final number of molecules produced in 30 cycles of the PCR process is:

A) 22
B) 210
C) 220
D) 230

Question 18

Multiplex PCR is limited by primer-primer interference.

Question 19

The name of the product of PCR is an:

A) amp.
B) amplicon.
C) anneal.
D) extension.

Question 20

MATCHING
Match the technique with an appropriate description (use each answer only once).

Strand displacement amplification

A)This amplifies target nucleic acid without the use of a thermocycler.A double-stranded DNA fragment is created and becomes the target for exponential amplification.
B)An isothermal assay that targets either DNA or RNA but generates RNA as its amplified product.
C)Oligonucleotide pairs hybridize to target sequences within the gene or the cryptic plasmid.
D)Only RNA is targeted for amplification.

Question 21

MATCHING
Match the technique with an appropriate description (use each answer only once).

Ligase chain reaction nucleic acid amplification

A)This amplifies target nucleic acid without the use of a thermocycler.A double-stranded DNA fragment is created and becomes the target for exponential amplification.
B)An isothermal assay that targets either DNA or RNA but generates RNA as its amplified product.
C)Oligonucleotide pairs hybridize to target sequences within the gene or the cryptic plasmid.
D)Only RNA is targeted for amplification.

Question 22

Match the hybridization technique with the correct description.

Reverse dot blot

A)Hybridization occurs only if the patient's DNA contains base sequences that are 100% complementary to those of the probe.
B)Both the target nucleic acid and labeled probe interact in solution.
C)This detects single-base mutations using allele-specific oligonucleotides (ASOs).

Question 23

Match the blotting protocol with the clinical application.

Northern blot

A)RNA
B)DNA
C)Use of labeled antibodies specific for the protein of interest

Question 24

Match the hybridization technique with the correct description.

Dot blot

A)Hybridization occurs only if the patient's DNA contains base sequences that are 100% complementary to those of the probe.
B)Both the target nucleic acid and labeled probe interact in solution.
C)This detects single-base mutations using allele-specific oligonucleotides (ASOs).

Question 25

Match the molecular technique with the appropriate microorganism identification.
Transcription-mediated amplification (TMA)

A)Cytomegalovirus (CMV)
B)Mycobacterium tuberculosis

Question 26

Match the molecular technique with the appropriate microorganism identification.
Nucleic acid sequence-based amplification (NASBA)

A)Cytomegalovirus (CMV)
B)Mycobacterium tuberculosis

Question 27

Match the hybridization technique with the correct description.

Liquid-phase hybridization

A)Hybridization occurs only if the patient's DNA contains base sequences that are 100% complementary to those of the probe.
B)Both the target nucleic acid and labeled probe interact in solution.
C)This detects single-base mutations using allele-specific oligonucleotides (ASOs).

Question 28

Match the blotting protocol with the clinical application.

Western blot

A)RNA
B)DNA
C)Use of labeled antibodies specific for the protein of interest

Question 29

Match the blotting protocol with the clinical application.

Southern blot

A)RNA
B)DNA
C)Use of labeled antibodies specific for the protein of interest