Question 1

Nested polymerase chain reaction (PCR) is very sensitive and specific because:

Accepted Answer

A) it uses more than one DNA polymerase to make three to four times the amount of PCR products than standard PCR. 
B) the assay itself basically serves as a form of internal control and ensures specificity. 
C) the assay will act as a form of external control, ensuring specificity and sensitivity. 
D) the fluorochrome that is attached to the primers ensures specificity of the nucleotides that are attached to the template. 
A) it uses more than one DNA polymerase to make three to four times the amount of PCR products than standard PCR. 
B) the assay itself basically serves as a form of internal control and ensures specificity. 
C) the assay will act as a form of external control, ensuring specificity and sensitivity. 
D) the fluorochrome that is attached to the primers ensures specificity of the nucleotides that are attached to the template.

Question 2

The basis for this method is a microscopic grouping of DNA molecules attached to a solid support mechanism.Silicon chips, glass, or plastic have been used as the solid surfaces.What method is this?

Accepted Answer

A) Multilocus enzymes electrophoresis 
B) Restriction fragment length polymorphism (RFLP) 
C) Polymerase chain reaction (PCR) 
D) DNA microarray 
A) Multilocus enzymes electrophoresis 
B) Restriction fragment length polymorphism (RFLP) 
C) Polymerase chain reaction (PCR) 
D) DNA microarray

Question 3

The goal of the primer annealing step is to:

Accepted Answer

A) hybridize oligonucleotide primers to the denatured, single-stranded target DNA strands. 
B) separate the target DNA so that the solution can be electrophoresed and a Northern blot performed. 
C) cut the native DNA into small pieces with a restriction enzyme. 
D) amplify exponentially over many cycles of these three reaction steps. 
A) hybridize oligonucleotide primers to the denatured, single-stranded target DNA strands. 
B) separate the target DNA so that the solution can be electrophoresed and a Northern blot performed. 
C) cut the native DNA into small pieces with a restriction enzyme. 
D) amplify exponentially over many cycles of these three reaction steps.

Question 4

Many hybridization assay conditions are based on the expectation that a probe:

Accepted Answer

A) will quickly react with the complementary target sequence. 
B) is not sterically hindered in the reaction. 
C) has exact complementarity to the target nucleic acid. 
D) will not require any oligonucleotides to attach to the target. 
A) will quickly react with the complementary target sequence. 
B) is not sterically hindered in the reaction. 
C) has exact complementarity to the target nucleic acid. 
D) will not require any oligonucleotides to attach to the target.

Question 5

Which of the following compounds are used for labeling probes?

Accepted Answer

A) Agar 
B) Fluorescein 
C) Digitoxin 
D) Horseradish peroxidase 
A) Agar 
B) Fluorescein 
C) Digitoxin 
D) Horseradish peroxidase

Question 6

How do higher probe concentrations affect a given hybridization reaction?

Accepted Answer

A) Increase the temperature. 
B) Lower the reaction time. 
C) Lower the ionic strength. 
D) Increase the pH. 
A) Increase the temperature. 
B) Lower the reaction time. 
C) Lower the ionic strength. 
D) Increase the pH.

Question 7

PCR requires all of the following components except:

Accepted Answer

A) DNA helicase. 
B) DNA polymerase. 
C) oligonucleotides (primers). 
D) deoxynucleotide triphosphates. 
A) DNA helicase. 
B) DNA polymerase. 
C) oligonucleotides (primers). 
D) deoxynucleotide triphosphates.

Question 8

The target nucleic acid strand is the:

Accepted Answer

A) one to which the extra nucleic acid attaches. 
B) DNA or RNA sequence unique to the organism of interest. 
C) strand that is used as the oligonucleotide primer. 
D) one that is labeled with a chromogen. 
A) one to which the extra nucleic acid attaches. 
B) DNA or RNA sequence unique to the organism of interest. 
C) strand that is used as the oligonucleotide primer. 
D) one that is labeled with a chromogen.

Question 9

What is the purpose of the primer extension?

Accepted Answer

A) To cut the native DNA into small pieces with a restriction enzyme 
B) To hybridize the oligonucleotide primers to the single stranded DNA pieces 
C) To produce PCR products 
D) To activate the DNA polymerase to form hybrids with the oligonucleotide primers 
A) To cut the native DNA into small pieces with a restriction enzyme 
B) To hybridize the oligonucleotide primers to the single stranded DNA pieces 
C) To produce PCR products 
D) To activate the DNA polymerase to form hybrids with the oligonucleotide primers

Question 10

What is the fluorescence resonance energy transfer (FRET) methodology?

Accepted Answer

A) The transfer of energy is made from a donor dye molecule to an acceptor dye molecule. 
B) A fluorescent probe is mixed with labeled primers and a sandwich occurs where the fluorescent molecule is activated to fluorescent. 
C) A light shines on the fluorescent probe label and it fluoresces. 
D) A chromogenic substrate is mixed into solution, and when it combines with the Taq DNA polymerase, a fluorophore is formed and it fluoresces. 
A) The transfer of energy is made from a donor dye molecule to an acceptor dye molecule. 
B) A fluorescent probe is mixed with labeled primers and a sandwich occurs where the fluorescent molecule is activated to fluorescent. 
C) A light shines on the fluorescent probe label and it fluoresces. 
D) A chromogenic substrate is mixed into solution, and when it combines with the Taq DNA polymerase, a fluorophore is formed and it fluoresces.

Exam 11: Applications of Molecular Diagnostics

Nested polymerase chain reaction (PCR) is very sensitive and specific because:

The basis for this method is a microscopic grouping of DNA molecules attached to a solid support mechanism.Silicon chips, glass, or plastic have been used as the solid surfaces.What method is this?

The goal of the primer annealing step is to:

Many hybridization assay conditions are based on the expectation that a probe:

Which of the following compounds are used for labeling probes?

How do higher probe concentrations affect a given hybridization reaction?

PCR requires all of the following components except:

The target nucleic acid strand is the:

What is the purpose of the primer extension?

What is the fluorescence resonance energy transfer (FRET) methodology?

Bacterial Cell Structure, Physiology, Metabolism, and Genetics

Host-Parasite Interaction

The Laboratory Role in Infection Control

Control of Microorganisms

Performance Improvement in the Microbiology Laboratory

Specimen Collection and Processing

Microscopic Examination of Materials From Infected Sites

Use of Colony Morphology for the Presumptive Identification of Microorganisms

Biochemical Identification of Gram-Negative Bacteria

Immunodiagnosis of Infectious Diseases

Antimicrobial Agent Mechanisms of Action and Resistance

Antimicrobial Susceptibility Testing

Staphylococci

Streptococcus, Enterococcus, and Other Catalase-Negative, Gram-Positive Cocci

Aerobic Gram-Positive Bacilli

Neisseria Species and Moraxella Catarrhalis

Haemophilus and Other Fastidious Gram-Negative Bacilli

Enterobacteriaceae

Vibrio, Aeromonas, Plesiomonas, and Campylobacter Species

Nonfermenting and Miscellaneous Gram-Negative Bacilli

Anaerobes of Clinical Importance

The Spirochetes

Chlamydia, Rickettsia and Similar Organisms

Mycoplasma and Ureaplasma

Mycobacterium Tuberculosis and Nontuberculous Mycobacteria

Medically Significant Fungi

Diagnostic Parasitology

Clinical Virology

Agents of Bioterror and Forensic Microbiology

Biofilms: Architects of Disease

Upper and Lower Respiratory Tract Infections

Skin and Soft Tissue Infections

Gastrointestinal Infections and Food Poisoning

Infections of the Central Nervous System

Bacteremia and Sepsis

Urinary Tract Infections

Genital Infections and Sexually Transmitted Diseases

Infections in Special Populations

Zoonotic Diseases

Ocular Infections

Filters