Molecular Diagnostics
Exam Answer Key
Course Introduction
Molecular Diagnostics explores the principles and techniques used to detect and analyze nucleic acids and proteins at the molecular level for disease diagnosis, monitoring, and therapy selection. The course covers key methodologies such as PCR, real-time PCR, DNA sequencing, microarrays, and next-generation sequencing, emphasizing their clinical applications in identifying infectious agents, genetic disorders, and cancer markers. Students learn about sample collection, assay development, validation, and quality control, as well as ethical and regulatory considerations. Through lectures, case studies, and laboratory exercises, the course prepares students for roles in clinical laboratories, research, and biotechnology industries.
Recommended Textbook
Molecular Diagnostics Fundamentals Methods and Clinical Applications 2nd Edition by Lela Buckingham
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16 Chapters
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Chapter 1: DNA
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Sample Questions
Q1) In a DNA molecule, one end has a free hydroxyl group and one end has a free
A) phosphate group
B) hydroxyl group
C) amino group
D) carboxyl group
Answer: A
Q2) What reactions are catalyzed by DNA methyltransferase?
A) Addition of methyl groups to nitrogen bases
B) Addition of methyl groups to ribose sugars
C) Addition of methyl groups to proteins
D) Removal of methyl groups from DNA
Answer: A
Q3) The purpose of ligase is to:
A) Synthesize DNA
B) Cut DNA within the double helix
C) Chew DNA from the ends
D) Paste two ends of DNA together
Answer: D
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Page 3
Chapter 2: RNA
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Sample Questions
Q1) Synthesis of DNA guided by a RNA template is:
A) Translation
B) DNA replication
C) Transcription
D) Reverse transcription
Answer: D
Q2) If the following oligonucleotide of double-stranded DNA were transcribed, what would be the sequence of the RNA? 5'TGCTAGCTA3' 3'ACGATCGAT5'
A) 5'UGCUAGCUA3'
B) 5'ACGAUCGAU3'
C) 3'ACGATCGAT5'
D) 3'ACGAUCGAU5'
Answer: A
Q3) Which of the following enzymes performs transcription in bacteria?
A) RNA-dependent DNA polymerase
B) DNA-dependent RNA polymerase
C) DNA-dependent DNA polymerase
D) RNA-dependent RNA polymerase
Answer: B
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Chapter 3: Proteins
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Sample Questions
Q1) Specialized proteins that bind to the large ribosomal subunit and protect the hydrophobic regions of growing polypeptides are called:
A) Aminoacyl tRNA synthetases
B) Molecular chaperones
C) Metalloproteins
D) Peptidyl transferases
Answer: B
Q2) Denaturation of a protein results in what effect?
A) Activation of protein activity
B) Loss of tertiary structure and protein function
C) None
D) Alteration of the amino acid sequence
Answer: B
Q3) The start codon for synthesis of most proteins is:
A) ACC
B) UGA
C) AUG
D) UAA
Answer: C
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Page 5
Chapter 4: Nucleic Acid Extraction Methods
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Sample Questions
Q1) In organic isolation of RNA procedures, guanidinium isothiocyanate is added to:
A) Degrade DNA
B) Denature all proteins
C) Inhibit RNAses
D) Separate RNA from DNA
Q2) In organic DNA extraction methods, the DNA is found in which fraction?
A) Upper aqueous
B) Lower aqueous
C) Upper hydrophobic
D) Lower hydrophobic
Q3) Isolated DNA has an absorbance at 260 nm of 0.268 and an absorbance at 280 nm of 0.191. What is the 260 nm/280 nm ratio?
A) 1:2
B) 1:4
C) 1:6
D) 1:8
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Chapter 5: Resolution and Detection of Nucleic Acids
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Sample Questions
Q1) To increase the density of a sample relative to the density of the buffer prior to loading the sample into a gel, which of the following is added to the sample?
A) Glycerol
B) Ethidium bromide
C) Formaldehyde
D) Tris acetate EDTA
Q2) A very limited amount of nucleic acid, 500-1500 bp in size, is to be analyzed in a short time (same day) with the results available immediately. Which of the following electrophoresis procedures will satisfy those conditions?
A) Capillary electrophoresis
B) Pulse-field gel electrophoresis
C) Field inversion gel electrophoresis
D) Contour-clamped homogeneous electric field
Q3) Which is an important property of gel running buffers?
A) Solubility at low temperature
B) Ability to maintain constant pH
C) Capacity to heat up quickly
D) High ultraviolet light absorption
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Chapter 6: Analysis and Characterization of Nucleic Acids and Proteins
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Sample Questions
Q1) A Southern blot procedure has been performed, but the molecular weight markers are the only bands seen on the gel. Apparently, the probe did not bind to its target on the membrane. How might the stringency be adjusted so that the probe will bind?
A) Decrease the incubation temperature.
B) Decrease the salt concentration.
C) Increase the formamide concentration.
D) Increase the incubation temperature.
Q2) A circular molecule with one restriction enzyme recognition site will yield how many fragments when digested by the restriction enzyme?
A) 1
B) 2
C) 3
D) 4
Q3) Which of the following solutions has the highest stringency?
A) 0% formamide
B) 10% formamide
C) high-salt buffer
D) 50% formamide
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Chapter 7: Nucleic Acid Amplification
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Sample Questions
Q1) A DNA sequence with high GC content is more likely to have what property that interferes with PCR amplification?
A) Tendency to degrade
B) Enzyme inactivation
C) Lower Tm
D) Secondary structure
Q2) What is the advantage of a nested PCR procedure?
A) Nested PCR is less labor-intensive.
B) Nested PCR offers increased specificity and yield of product.
C) Nested PCR has a shorter reaction time than standard PCR.
D) Nested PCR is less expensive than standard PCR.
Q3) Which of the following PCR modifications was developed to allow for the amplification of an RNA template?
A) Multiplex
B) Nested
C) Reverse transcriptase
D) Real-time
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9
Chapter 8: Chromosomal Structure and Chromosomal
Mutations
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Sample Questions
Q1) Human chromosomes 21 and 22 are described as which of the following with regard to the location of the centromere?
A) Telocentric
B) Holocentric
C) Metacentric
D) Acrocentric
Q2) Before its conversion to a microarray technique, CGH was performed by hybridizing labeled DNA to what support?
A) Immobilized PCR products
B) Normal chromosome spread
C) Nitrocellulose membrane
D) Agarose gel
Q3) Which of the following types of chromatin are open and actively involved in gene expression (transcription)?
A) Heterochromatin
B) Homochromatin
C) Euchromatin
D) Interchromatin
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Chapter 9: Gene Mutations
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Sample Questions
Q1) Which of the following methods has the highest sensitivity, specificity, and accuracy as far as detecting DNA mutations in clinical applications?
A) ELISA
B) High-density oligonucleotide array
C) PCR-RFLP
D) Single-strand conformation polymorphism
Q2) A reference segment of DNA has the following sequence: GCTCACCATGG. Insertion of AG occurs between positions 7 and 8 in a particular genetic disease. How is this mutation denoted by accepted nomenclature?
A) 7C>AG
B) 7_8insAG
C) 8delAinsAG
D) 7_8delAG
Q3) Which of the following methods can be used to screen for the presence of unknown mutations?
A) SSP-PCR
B) Allele-specific oligomer hybridization
C) High-density array
D) PCR-RFLP
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Chapter 10: DNA Sequencing
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Sample Questions
Q1) In bioinformatics, when a sequence is entered and compared with all other sequences in a database, which of the following is being performed?
A) Algorithm
B) Query
C) Motif
D) Annotation
Q2) Dideoxycytidine is added to one of four sequencing reactions. All of the DNA fragments produced in that reaction end with which of the following nucleotides?
A) dG
B) ddC
C) dC
D) ddG
Q3) The goal of the HapMap project is which of the following?
A) Find blocks of SNPs that are inherited together
B) Complete the human genome sequence
C) Compare human and primate genomes
D) Sequence all bacterial genomes
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Chapter 11: DNA Polymorphisms and Human Identification
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Sample Questions
Q1) Southern blot is used to detect which of the following polymorphisms?
A) Edited RNA
B) Alternative splicing
C) Restriction fragment length polymorphisms
D) Modified nucleotides
Q2) How are Y alleles inherited?
A) As recombined combinations on two chromosomes
B) As a diploid genotype
C) As a haplotype on one chromosome
D) Maternally
Q3) Two peaks from different sources falling within a bin describe what relationship?
A) Different alleles
B) Identical alleles
C) Paternity
D) Exclusion
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13
Chapter 12: Detection and Identification of Microorganisms
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Sample Questions
Q1) What will be observed in a PCR test of a true negative sample for the sample, positive control, negative control, and amplification control, respectively?
A) Negative, positive, negative, positive
B) Negative, positive, negative, negative
C) Negative, negative, negative, negative
D) Negative, negative, negative, positive
Q2) A positive control is added (spiked) into a test specimen. The test results of the specimen alone and the spiked mixture are negative. What does this most likely indicate?
A) The specimen is negative.
B) The specimen is positive.
C) The specimen contains an inhibitor.
D) The positive control contains an inhibitor.
Q3) Which is a flow cytometry test?
A) Hybrid capture
B) NASBA
C) TMA
D) HPV OncoTect
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14
Chapter 13: Molecular Detection of Inherited Diseases
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Sample Questions
Q1) Mitochondrial mutations primarily affect which of the following tissues?
A) Skin
B) Bone
C) Muscle
D) Lung
Q2) Genetic analysis of four generations of a family reveals that about 50% of the males in the family are affected by a mutation and about 50% of the females are unaffected carriers of the mutation that affects the males. What type of inheritance pattern is seen in this family?
A) Autosomal-dominant
B) Autosomal-recessive
C) X-linked recessive
D) Y-linked recessive
Q3) Mutation in this gene product that catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate can lead to cardiovascular disease and complications in pregnancy.
A) TP53
B) MTHFR
C) F5
D) HLA-H
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Chapter 14: Molecular Oncology
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Sample Questions
Q1) What would be observed in the event of loss of heterozygosity?
A) Two alleles at a heterozygous locus
B) One allele at a homozygous locus
C) One allele at a heterozygous locus
D) Two alleles at a homozygous locus
Q2) A patient is suspected of having follicular lymphoma, and his physician would like additional laboratory tests performed to confirm or rule out the diagnosis. Which of the following tests should be performed?
A) T-cell receptor gene rearrangement clonality
B) t(9;22) translocation
C) t(14;18) translocation
D) t(11;14) translocation
Q3) Gene expression patterns are now used to characterize tumor types. What method is appropriate for analysis of expression of many genes simultaneously?
A) Northern blot
B) GE arrays
C) CGH arrays
D) Slot blots
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16
Chapter 15: DNA-Based Tissue Typing
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Sample Questions
Q1) What does the term HLA-DQ designate?
A) HLA gene region, locus Q, sub-region D
B) The entire MHC gene region, locus D
C) HLA gene region, locus D, sub-region Q
D) The entire HLA-D locus
Q2) HLA is an abbreviation for what term?
A) Histocompatability locus array
B) Human leukocyte antibody
C) Human leukocyte antigen
D) Hepatic lipid antigen
Q3) Bone marrow is going to be transplanted from a man into his sister. Which of the following laboratory procedures should be performed prior to this transplant to determine the HLA alleles of the donor and recipient at the highest resolution?
A) SBT
B) CDC
C) PCR-SSP
D) PCR-SSOP
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Chapter 16: Quality Assurance and Quality Control in the Molecular Laboratory
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Sample Questions
Q1) The ability of a molecular-based assay to detect only the analyte and not other targets is called which of the following?
A) Clinical specificity
B) Clinical sensitivity
C) Analytic sensitivity
D) Analytic specificity
Q2) Which test method is best for a fixed tissue specimen?
A) Long-range PCR
B) Southern blot
C) Standard PCR
D) PFGE
Q3) A blood specimen is received in the laboratory for a PCR assay to detect cytomegalovirus (a DNA virus) on a Wednesday night. The PCR assay will not be performed until the next Monday morning. How should this specimen be stored to maintain the integrity of the nucleic acid prior to analysis?
A) Lyze RBCs, centrifuge, and store at 22<sup>o</sup>C
B) Lyze RBCs, centrifuge, and store at -20<sup>o</sup>
C) Whole blood stored at 22<sup>o</sup>-25<sup>o</sup>C
D) Whole blood stored at 35<sup>o</sup>-37<sup>o</sup>C
Page 18
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