Chapter 11: Molecular Techniques Flashcards

1
Q

What represents a significant advancement in clinical microbiology?

A

Molecular Diagnostics

Considered the fastest growing section in many clinical laboratories.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

What do molecular diagnostics provide to clinicians?

A

Rapid answers for treatment options

Saves valuable time in cases of life-threatening infections.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

When are molecular techniques particularly useful?

A

When traditional methods are not practical or available.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Which organisms cannot be routinely cultivated?

A

Viruses and Chlamydia

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What is required for the growth of Mycobacterium tuberculosis?

A

A long incubation period

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What are genetic probes?

A

Short, specific sequences of single-stranded DNA or RNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What do genetic probes identify within nucleic acid?

A

One or more sequences of interest

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

What does the degree of hybridization depend on?

A

Base sequence homology and reaction conditions

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What are the conditions that affect stringency?

A

Length of probe, probe concentration, degree of complementarity, salt concentration, temperature, pH, concentration of destabilizing agents

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What determines the detection of hybridization?

A

Type of label

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What is the most common method for detection in clinical microbiology?

A

Chemiluminescence

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

List the formats of nucleic acid probes.

A
  • Liquid phase
  • Solid phase
  • Southern blot
  • Northern blot
  • In situ hybridization
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

What is signal amplification in nucleic acid amplification methods?

A

Increased concentration of labeled molecule without increasing probe or target concentration.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Branched DNA amplification:

A

Solid phase sandwich hybridization that uses multiple sets of synthetic probes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Hybris capture assays:

A

Solution hybridization antibody capture

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Target amplification:

A

– Enzymes produce copies of target nucleic acid which are detected by primers that bind to complementary strands.

– Produces millions of copies of targeted sequence which act as templates for subsequent amplification cycles.

– Polymerase chain reaction (PCR)

17
Q

What is the purpose of Polymerase Chain Reaction (PCR)?

A

To produce millions of copies of targeted nucleic acid sequence

18
Q

What is the first step in the Polymerase Chain Reaction?

A

Denature nucleic acid into single stranded DNA

19
Q

What is Reverse Transcriptase PCR (RT-PCR) used for?

A

To detect RNA viruses

20
Q

What does Nucleic Acid Sequence-Based Amplification (NASBA) require?

A

Three enzymes: AMV reverse transcriptase, Ribonuclease H, T7 RNA polymerase

Target could be DNA or RNA most often RNA viruses

21
Q

What is Transcription-Mediated Amplification (TMA) developed for?

A

Detection of C. trachomatis and N. gonorrhoeae

Targets rRNA

22
Q

What is strain typing and identification based on?

A

DNA or genetic fingerprinting and genetic relatedness

23
Q

What are Amplification Independent Methods?

A
  • Restriction Fragment Length Polymorphisms (RFLPs)
  • Southern blotting
  • Plasmid profile analysis
  • Multilocus enzyme electrophoresis (MLEE)
24
Q

List some Amplification Dependent Methods.

A
  • Random amplified polymorphic DNA
  • Repetitive palindromic extragenic elements PCR
  • Multilocus sequence typing
25
What is a DNA microarray?
Grouping at the micron level of DNA molecules attached to a solid support
26
What are DNA chips used for?
Investigation of gene expression from an entire organism
27
What is the purpose of nanoarrays?
To analyze proteins and gene expression at nanometer spatial resolutions
28
What is Sanger Sequencing?
Automated DNA sequencing method
29
What does MALDI-TOF stand for?
Matrix-Assisted Laser Desorption–Ionization Time of Flight
30
What is the primary use of MALDI-TOF?
Rapid identification of microorganisms
31
What are the three basic nucleic acid molecular diagnostics applications?
* Nucleic acid hybridization assays * Nucleic acid amplification techniques * Strain typing techniques
32
Nucleic acid hybridization assays – use
labeled probes
33
Nucleic acid amplification techniques – use
PCR and varients