Clin Path Exam 3 Review Flashcards
1.single stranded building blocks
1 sugar 1 phosphate 1 base
2.either splicing
post transcriptional modification
3.Cleaves or cut the DNA molecules at the recognition site
a. DNA ligases
b. Endocuclease
c. Exonuclease
(Restriction) Endonuclease
Bacterial endonucleases that recognize specific short DNA base pair sequences and cleave the DNA molecule only at the recognition site
- Which of the following is the correct sequence for DNA?
Initiation -> Elongation -> Termination
Steps of DNA synthesis in order
a. chain
b. linked together by phosphodiester
c. nucleotides
d. stripped to two phosphate groups
Nucleotides → stripped to two phosphate groups → linked together by phosphodiester bonds → chain
- Sections of DNA that specify amino acid sequences of proteins
Codons
- Which faster and more specific test which will determine a gene and specific mutation or presence of particular CHROMOSOMAL REARRANGEMENT?
FISH
fluorescent in situ hybridization
enables detection of gene rearrangements and gene deletions in a number of diseases, especially in cancers
- Short, circular dsDNA segments that can be used as vectors in propagating desired segments in bacteria
Plasmid
- Detection of specific genetic information within a
MORPHOLOGIC CONTEXT?
In situ hybridization
- Amplify DNA
PCR
- Used to know the amounts of DNA and RNA in a sequence
a. Helicase - Dependent Amplification
b. Loop - Mediated Amplification
c. nucleic acid sequence-based amplification (NASBA)
d. Strand - Displacement Amplification
e. transcription-mediated amplification (TMA)
d. Stranded Displacement
- Amplifies RNA targets followed by cDNA amplification
a. Digital Polymerase Chain Reaction
b. End-Point Quantitative Polymerase Chain Reaction
c. Multiple Polymerase Chain Reactions
d. Nested Polymerase Chain Reaction
e. Rapid-Cycle Polymerase Chain Reaction
f. Real-Time (Homogenous, Kinetic) Polymerase Chain Reaction
g. Reverse-Transcriptase Polymerase Chain Reaction
reverse transcriptase polymerase chain reaction
- Uses two amplification primers
a. Digital Polymerase Chain Reaction
b. End-Point Quantitative Polymerase Chain Reaction
c. Multiple Polymerase Chain Reactions
d. Nested Polymerase Chain Reaction
e. Rapid-Cycle Polymerase Chain Reaction
f. Real-Time (Homogenous, Kinetic) Polymerase Chain Reaction
g. Reverse-Transcriptase Polymerase Chain Reaction
Nested polymerase chain reaction
mRNA - Messenger RNA:
Encodes amino acid sequence of a polypeptide.
tRNA - Transfer RNA:
Brings amino acids to ribosomes during translation
rRNA - Ribosomal RNA:
With ribosomal proteins, makes up the ribosomes, the organelles that translate the mRNA.
snRNA - Small nuclear RNA:
With proteins, forms complexes that are used in RNA processing in eukaryotes. (Not found in prokaryotes.)
- Two or more primer sets for amplification of different targets included in the same reaction mixture
a. Digital Polymerase Chain Reaction
b. End-Point Quantitative Polymerase Chain Reaction
c. Multiple Polymerase Chain Reactions
d. Nested Polymerase Chain Reaction
e. Rapid-Cycle Polymerase Chain Reaction
f. Real-Time (Homogenous, Kinetic) Polymerase Chain Reaction
g. Reverse-Transcriptase Polymerase Chain Reaction
c. Multiple Polymerase Chain Reactions
Multiplex PCR
- Target amplification and detection steps occur simultaneously in the same tube
Real time PCR
Isothermal amplification technology
a. End-Point Quantitative Polymerase Chain Reaction
b. Loop - Mediated Amplification (LAMP)
c. Multiple Polymerase Chain Reactions
d. Real-Time (Homogenous, Kinetic) Polymerase Chain Reaction
Loop - Mediated Amplification (LAMP)
also:
- transcription-mediated amplification (TMA)
- nucleic acid sequence-based amplification (NASBA)
- Strand - Displacement Amplification
- Helicase - Dependent Amplification
- RFLP (Restriction Fragment Length Polymorphism)
Southern Blotting - DNA sample
Stop codons
UAG, UGA, or UAA
Polymerases
a. catalyzes the formation of
phosphodiester bonds during synthesis
b. hydrolyzes phosphodiester bonds
c. found only in bacteria that functions to destroy foreign DNA
a. catalyzes the formation of
phosphodiester bonds during synthesis
Nucleases
a. catalyzes the formation of
phosphodiester bonds during synthesis
b. hydrolyzes phosphodiester bonds
c. found only in bacteria that functions to destroy foreign DNA
b. hydrolyzes phosphodiester bonds
Restriction endonuclease
a. catalyzes the formation of
phosphodiester bonds during synthesis
b. hydrolyzes phosphodiester bonds
c. found only in bacteria that functions to destroy foreign DNA
c. found only in bacteria that functions to destroy foreign DNA
Assay utilizing Formaldehyde gel
Northern Blot - RNA sample
Assay utilizing Polyacrylamide gel
Western blot - proteins
- Cytokine genes characterization.
a. cDNA MICROARRAYS
b. OLIGONUCLEOTIDE MICROARRAYS
c. SEQUENCING ARRAYS
b. OLIGONUCLEOTIDE MICROARRAYS
Application: Gene expression profiling and DNA sequencing
- DNA sequencing-
microarray
Offer a platform for quantifying the expression of thousands of genes at the same time
Steps of PCR
Denaturation -> Annealing -> Extension
Which will melt at higher temperature?
a. A-T bond
b. G-C bond
b. G-C bond (has 3 hydrogen bonds)
A-T bonds have 2 hydrogen bonds
Melting point is dependent on
- length
- G:C content
- amount of mismatch
blotting techniques are used for
specific identification of desired DNA or RNA fragments from thousands of cells
What is blotting?
process of immobilization of sample nucleic acids on solid support
blotted nucleic acids are used as targets for
hybridization experiments
Southern blot used for
DNA
Northern blot used for
RNA
Western blot used for
proteins
FLUORESCENCE IN SITU HYBRIDIZATION
o Combination of molecular and cytogenic techniques
o Molecular probe binds to the chromosome instead of a dye
o Quicker, more specific and allows the use of multiple probes in a single hybridization
o Most common goal is to determine whether a gene, a specific mutation, or a particular chromosomal rearrangement is present
Most common pattern of inheritance
Autosomal Dominant (AD)
- at least 1 parent is affected
- males and females affected with equal frequency
Penetrance:
refers to the proportion of persons with the mutation who manifest a phenotypic abnormality
Expressivity:
refers to the severity and range of manifestation
- Epigenetic mechanism is affected by the following factors except:
Genotype
Karyotype
Phenotype
Genotype
Result of abnormal division of centromere
Isochromosome
Highly ordered structure composed of DNA and
proteins that carry genetic info
chromosome
- Appearance of an individual from an interaction of environment and genotype
Phenotype
- Strep pneumoniae and H. influenzae
B cell and complement defects
- Recurrent viral fungal and opportunistic infection has a deficiency in:
a. B-Cell lymphocyte
b. Neutrophils
c. T-Cell lymphocyte
T-Cell lymphocyte
- A pt. report hx of infection with catalase positive
bacteria such as staph. -
phagocytic
centrioles moves to opposite poles
a. Anaphase
b. Meiosis
c. Metaphase
d. Prophase
d. Prophase
nuclear envelope disappear and chromosome condensation
a. Anaphase
b. Meiosis
c. Metaphase
d. Prophase
c. Metaphase
separation of sister chromatids
a. Anaphase
b. Meiosis
c. Metaphase
d. Prophase
a. Anaphase
occurs only in germ cells
a. Anaphase
b. Meiosis
c. Metaphase
d. Prophase
b. Meiosis
S phase
- DNA replication
- semi conservative
- DNA helicase and DNA topoisomerase unwinds the double helix
- occurs at multiple sites
Polymorphism: (is/is not) deleterious and present at least in at least 1% o population
Polymorphism: not deleterious and present at least in at least 1% o population
genetic change often with a potential deleterious effect
Mutation: genetic change often with a potential deleterious effect
- Signs and symptoms of primary immunodeficiency.
> 2 episode of pneumonia per year
30.(non-verbatim) 11-month old male. With SMALL OR ABSENT PERIPHERAL LYMPH NODES.
X-linked agammaglobulinemia
- A 5-month-old male has history of bloody diarrhea and recurrent bacterial infections. On physical examination, you noticed bruises and EXZEMATOUS SKIN RASHES. What is the most likely diagnosis?
Wiskott-Aldrich syndrome
- Primary investigation of the immune system?
A. CBC B. Radiograph C. Quantitative Immunoglobulins (IgG, IgM, IgA) D. Pulmonary function testing E. All of the above
E. All of the above
Tumor Marker
CA 125
Ovarian carcinoma
Tumor Marker
CA 19-9
Pancreatic carcinoma
Tumor Marker
CA 15-3
Breast carcinoma
Tumor Marker
CA 72-4
Gastric carcinoma
Tumor Marker
HER2/neu
Breast carcinoma
B cell and complement defects
- streptococcus pneumoniae
- haemophilius influenza
- mycoplasma pneumoniae
- neisseria meningitis
Phagocytic defects
- Pseudomonas aeruginosa
- Klebisella pneumoniae
- Serratia marcescens aspergillus
T cell defects
- Pneumocystis jiroveci
- Mycobacteria avium complex
- Cytomegalovirus
- Listeria monocytogenesis
- Herpes virus
Staphylococcus aureus is a deficiency in:
a. B cell and complement defects
b. T cell defects
c. Phagocytic defects
a. B cell and complement defects
c. Phagocytic defects
Candida is a deficiency in:
a. B cell and complement defects
b. T cell defects
c. Phagocytic defects
b. T cell defects
c. Phagocytic defects
Giardia Crytosporidia is a deficiency in:
a. B cell and complement defects
b. T cell defects
c. Phagocytic defects
a. B cell and complement defects
b. T cell defects
Signs and symptoms of primary immunodeficiency
- > 2 episode of pneumonia per year
- > 2 sinus infection/year
- > 8 new ear infections/year
- > 1 episode severe infection/year
- Review IV antibodies to resolve infections
- Family history
Secondary investigation of the immune system
- titers for vaccine administered
- IgG subclass analysis
- Lymphocyte enumeration panel
- Complement level
- Skin testing
- Mononuclear cell proliferation studies
- NBT
Staphylococcus spp. defect
phagocytic defects
Patients with immunoglobulin deficiencies typically report an increased number of (bacterial/viral) infections
Patients with immunoglobulin deficiencies typically report an increased number of bacterial infections
A 5-month-old male has history of bloody diarrhea and recurrent bacterial infections. On physical examination, you noticed NORMAL AND ENLARGED PERIPHERAL LYMPH NODES
- chronic granulomatous disease (CGD)
* common variable immunodeficiency
Streptococcus pneumoniae or Neisseria spp. infections defect
Complement deficiencies
Repeated viral, fungal, or other opportunistic infections are associated with
T cell immunodeficiencies
- The most common and accurate method of provocative challenge of hypersensitivity reactions.
Skin Test
- Laboratory test for cytotoxic rxn:
Direct coombs test
Hypersensitivity mediated by cell components (T-lymphocytes)
a. Type I
b. Type II
c. Type III
d. Type IV
d. Type IV hypersensitivity mediated by cell components (T-lymphocytes)
Types 1, 2, and 3- mediated by humoral components (antibodies and complement)
Match
Type 1:
Type 2:
Type 3:
Type 4:
a. Cell-mediated hypersensitivity reactions
b. Cytotoxic reaction
c. Immediate hypersensitivity reaction
d. Immune complex-mediated reactions
Type 1: Immediate hypersensitivity reaction.
Type 2: Cytotoxic reaction
Type 3: Immune complex-mediated reactions
Type 4: Cell-mediated hypersensitivity reactions
ACID
Anaphylaxis, Cytotoxic, Immune Complex, Delayed Type
Laboratory tests for anaphylaxis reaction - 2
- skin test
2. In vitro IgE assay (radioallergosorbent test- RAST)
Laboratory test for cytotoxic reaction - 2
- Direct Coombs’ test
2. Test for anti-thyroid peroxidase antibodies
- Laboratory test for immune complex mediated reaction
ANA
ANC
ANA
- Laboratory test for cell mediated reaction - 2
- Tissue biopsy
2. Skin test
Laboratory test for immune complex reaction - 4
- Erythrocyte sedimentation rate (ESR)
- Anti-nuclear Ab (ANA)
- Anti-streptolysin O (ASO)
- Histopath
- Clinical situations involving hypersensitivity reactions
A. Mycoplasma tuberculosis
B. Brucellosis
C. Chagas’ disease
D. AOTA
D. AOTA
- Clinical situation involving a post streptococcal glomerulonephritis
a. Type I
b. Type II
c. Type III
d. Type IV
c. Type III Hypersensitivity
- Clinical situations involving cytotoxic reactions
A. Autoimmune hemolytic anemia B. Autoimmune thrombocytopenic purpura C. Myasthenia gravis D. All of the above
D. All of the above
- Test for phagocytic activity
NBT
Test of cellular T-cell immunity
- 12
- CBC with absolute lymphocyte count
- Delayed hypersensitivity skin test to recall Ag
- Quantification of T-cell and T-cell subsets by flow cytometry
- Enzymatic tests: ADA (adenosine deaminase) and PNP (purine nucleoside phosphorylase)
- Cytokine receptor expression: IFNyR, IL-2A, gamma chain, IL-12R
- Tests of immunoregulation and immunoglobulin
synthesis. - lymphocyte blast transformation response to mitogens and antigens.
- mixed lymphocyte culture assays
- tests of lymphocyte-mediated toxicity
- molecular test for gene defects
- cytokine production assay by elisa: il2, il4, tgf-b, ifny, etc.
- Hiv testing as indicated.
Test of humoral (b-cell) immunity - 7
- Quantitation of serum igs
- Isotypes: igg, igm, iga, ige
- Igg and iga subclass levels
- Tests for functional abs
- Ab response after infection to respiratory viruses
- Enumeration and phenotyping of b cells in blood, surface igg, iga, igm, ige, igd
- Polyclonal b-cell induced ig production in vitro.
Test of phagocytic immunity - 6
- Cbc with absolute neutrophil count
- Nbt (nitoblue tetrazolium) or dihydrorhodamine flourescence test for respiratory burst activity
- Chemotaxis assay
- Serum ige level
- Phagocytosis and bacterial killing assay
- Flowcytometry for leukocyte adhesion molecules cd11/18.
Test of complement immunity
• (ch50 for primary complement deficiency)
• 2. Rabbit rbc screening for alternative pathway hemolytic
activity.
- Serum c2, c3, c4, c5 and factor b levels
- Individual component assay
- C1 esterase inhibitor concentrations
- Test for Humoral B cells
Quantitative Immunoglobulin
Functional abs
- Confirmatory test for inflammation
ESR and C-reactive protein
Tissue biopsy of the affected organ for histologic and immunohistochemical and immunofluoresecent study examinations
- Positive / Negative cutoffs
A. If the mean plus 2 standard
deviation is chosen as the cutoff, approx. 2.5% of the healthy population will test +
B. If the mean plus 3 standard deviation is chosen as the cutoff, approx. 0.15% of the health population should test +
C. A&B
D. Neither a & b
C. A&B
A. If the mean plus 2 standard
deviation is chosen as the cutoff, approx. 2.5% of the healthy population will test +
B. If the mean plus 3 standard deviation is chosen as the cutoff, approx. 0.15% of the health population should test +
- In Grave’s disease, following pharmacologic therapy, what is the relationship of the titer of TSI with likelihood of remission?
The titer of TSI are INVERSELY correlated with the likelihood of remission
- In nondiabetic individuals with ICA.
high ICA indicates subsequent development of type 1 diabetes (DIRECT CORRELATION)
- 43 year old, female patient has dry mouth
Sjögrens syndrome
- 15 year old, has intestinal malabsorption, thin lips, etc.
Centromeric pattern
Fine speckled nuclear pattern in indirect immunofluorescence assay
SJOGREN’S SYNDROME
Centromeric pattern in indirect immunfluorescence assay
SCLERODERMA
SJOGREN’S SYNDROME
A. Dry mouth and tongue.
B. Parotid enlargement.
C. Hypergammaglobulinemic purpura with photosensitivity
d. Fine speckled nuclear pattern in indirect immunofluorescence assay
SCLERODERMA
A. Intestinal malabsorption, radiologic study showing diffuse dilation and hypoperistaltic waves.
D. Collagen deposition brings thickness to the lingual frenulum.
E. Typical facial appearance in scleroderma: telangiectasias, lack of expression, thin
lips and nose.
F. Advanced sclerodactyly.
The dx of systemic rheumatic diseases depends on - 4
- clinical characteristics
- laboratory testing
- biopsy (occasionally)
- exclusion of other diseases
in systemic lupus erythematosus (SLE)
A. Scarring discoid lesions. B. Finger vasculitis. C. Vasculitis with ulcers. D. Subacute cutaneous geographic lesions.
RHEUMATOID ARTHRITIS
a. Subcutaneous nodules in elbow region, common in seropositive RA.
b. Episcleritis in RA.
c. Proximal interphalangeal joint synovitis of the third finger in juvenile RA.
d. Typical hand appearance in adult RA, with intrinsic muscle atrophy and cubital deviation of fingers
tests for RA
- APF, Anti-perinuclear factor;
- CCP, cyclic citrullinated peptide
- ELISA, enzyme-linked immunosorbent assay
- RF, rheumatoid factor
Laboratory tests for evaluating Autoimmune Diseases
Tissue biopsy of the affected organ for histologic and immunohistochemical and immunofluoresecent study examinations