Molecular Diagnostics Flashcards

1
Q

Southern Blotting

A

both probe and target nucleic acid are DNA

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2
Q

Northern Blotting

A

probe is single stranded DNA and target is mRNA

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3
Q

Western Blotting

A

measures amount of protein or antibody

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4
Q

Eastern Blotting

A

Detects post-translational modification on proteins

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5
Q

PCR

A

 Subjected to high temp. to denature or separate single stranded DNA
 Primers designed to complement sequences that flank each end of DNA in 3’-5’ direction
 Allowed to anneal
 Taq polymerase synthesizes copy of DNA by extending the primers on both ends
 Advantage: Very small amount of template DNA needed
 Disadvantage: Need to know the sequence of the flanking DNA for primer design, error prone

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6
Q

qPCR

A
  • used to quantify copy number of specific copy number of a specific gene in two or more samples in real time
  • this technique includes a probe which fluoresces only in presence of PCR product
  • used to detect levels of infectious agent
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7
Q

Restriction fragment length polymorphism (RFLP)

A

o Individual genomes differ by 1 in every 1000 base pairs
o Some of these occur in the recognition sequences for restriction enzymes
o DNA fingerprinting
o Used in forensic analysis, paternity testing and disease detection
Ex: detection of sickle cell disease and number of restriction sites

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8
Q

b. Variable number of tandem repeats (VNTR)

A

o Varies in individuals
o Regions isolated from genomic samples by flanking restriction sites or through PCR
o Useful in identification and severity of inherited diseases
o Huntington disease, Fragile X syndrome, Frederich Ataxia detection

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9
Q

i. Improving on insulin

A

o Normal human insulin has proline at position 28 and lysine at position 29 at C terminus of B chain
o Lispro reverse position of these aa (made it faster)
o Insulin aspart- proline 28 replaced by aspartic acid
o Mixed with normal insulin, provides longer range of glycemic control

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10
Q

 Abciximab

A

• Inhibits platelet aggregation

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11
Q

 Baciliximab

A

• Prevents rejection of transplanted kidney

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12
Q

 Cetuximab

A

• Treats metastatic colorectal cancer

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13
Q

 Infliximab

A

• Treats autoimmune diseases

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14
Q

 Retuximab

A

• Treats lymphomas, leukemias

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15
Q

Diagnosis of HIV

A

• Specific antibodies to HIV are produced in humans 4-6 weeks after infection
• Immobilized antigens in the wells with the biological samples
• Wash plate to remove unbound antibody and nonspecific antigen-antibody associations
• Add enzyme-linked antihuman IgG second antibody, wash, and add chromogenic substrates
• Development of color indicates presence of anti-HIV antibodies
- after sandwhich ELISA, then western blots to confirm

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