Amplified Nucleic Acic Tests

Question 1

Polymerase chain reaction (PCR)

Answer 1

Target amplification using DNA polymerase

Using of thermocycling - yes

Question 2

Ligase chain reaction (LCR)

Answer 2

Amplification of a short DNA probe using DNA ligase

Using of thermocycling - yes

Question 3

Transcription-based or transcription mediated amplification

Answer 3

Target amplification using reverse transcriptase and RNA polymerase

Using of thermocycling - No

Question 4

Strand displacement (SDA)

Answer 4

Target amplification using DNA polymerase that continuously displaces strands of DNA containing the target sequence

Using of thermocycling - No

Question 5

Branched DNA (bDNA)

Answer 5

Signal amplification using alkaline phosphatase

Question 6

Loop mediated (LAMP)

Answer 6

Target amplification of multiple DNA sequences in a loop pattern using DNA polymerase

Question 7

Nucleic acid sequence based (NASBA)

Answer 7

Target amplification using 3 enzymes

Question 8

Q-beta replicase

Answer 8

Probe amplification- the concentration of an RNA probe increases if the target is present

Question 9

Thermocycling

Answer 9

Uses specific instrumentation to cause variations in temp during an amplification procedure in order to produce copies, or amplifies, of the target interest so that detection can take place

Question 10

Amplicon

Answer 10

The product of an amplification reaction; amplicons are the outcome of what is to be detected

Question 11

Chemiluminescence

Answer 11

Release of light energy at the end of a chemical reaction that is detected by a luminometer. Uses a label as acridinium ester

Question 12

Electrophoresis

Answer 12

Movement in a matrix such as gel that is caused by an electrical field

Question 13

Enzyme (detection)

Answer 13

Uses enzyme and substrate principles to label the appropriate target or probe. Can be combined with fluorescence or dyes for detection

Question 14

Fluorescence

Answer 14

Molecules that emit light at a longer wavelength when excited at a shorter wavelength. Detection techniques include fluorescent staining of nucleic acids as well as fluorescent labeled probes that are measured in a fluorometer or with fluorescent polarization

Question 15

Radioactivity

Answer 15

Uses a labeling technique where the radioactive label is then measured in a scintillation counter. The earliest assays utilized radioactive decay

Question 16

Detection Techniques

Answer 16

Detection techniques can vary in both direct and amplified methodologies and can include labeling either the probe or the target molecule of interest:

• Chemiluminescence
• Electorphoresis
• Enzyme
• Fluorescence
• Radioactivity