-
How long are the DNA primers
16 to 20 bp
-
What are primers for PCR
Small portions of single stranded DNA
-
The primer sequence is complementary to the __ end to be amplified
3'
-
PCR cycles consists of
denaturation, annealing, extension of primed DNA.
-
What is annealing process
when primer binds to the 3' end
-
What enzyme is needed to add amino acids
DNA polymerase
-
DNA being amplified
amplicon
-
RT PCR area of interest is
RNA
-
three important applications of PCR are
amplification of DNA, identification of target sequence and synthesis of a labeled antisense probe
-
RT-PCR useful in identification of RNA viral agents
HIV and HCV
-
Real time PCR used _____ energy to quantitate specific DNA and identifiy ____ mutations
fluorescence resonance, point
-
Multiplex PCR numerous primers to amplify nucleic acids from
different targets
-
Dot and Reverse dot blot used in clinical lab for
detection of disorders in which sequences is mutated region is known (sickle, cystic fibrosis)
-
Dot blot and reverse dot blot capable of distiguishing _____ states of mutation
heterozygous and homozygous
-
Advantage of molecular testing is
faster turnaround times, smaller required sample volumes, increased specificity and sensitivity.
-
DNA and other biomolecules can be separated based on
charge, shape and size
-
DNA has a net charge ___ and will migrate towards
negative, positive (anode)
-
DNA sequencing is considered the ___ standard for which
gold standard, other methods are compared to.
-
Branched DNA an alternative quantitative test
uses signal amplification instead of target amplification
-
dot blot detects single base mutations based on
allele specific oligonucleotides (ASO)
-
Dot blot hybridization occurs when
labeled probe base sequence match perfectly the patient's
-
Reverse dot blot similiar except
ASO bound to filter
-
Solid support for Northern blot and Southern blot
nitrocellulose
-
Southern blot detects____ Northen blot detects___
DNA, mRNA
-
Southern blot can detect DNA sequences
RFLP
-
Western blot is used to separate
proteins using electrophoresis then transferred to membrane
-
PCR testing is useful in
forensic, genetic, disease diagnosis
-
For PCR reaction to occur, the clinician must provide which of the following?
primers, polymerase, DNA
-
The enzyme reverse transcriptase converts
mRNA to cDNA
-
DNA polymerases catalyzes
primer extension
-
Southern blot immunoassay
single stranded DNA is studied
-
Northern blot immunoassay
Mesenger RNA is studied
-
Western blot immunoassay
Called immunoblot detects antibodies
-
What techniques uses signal amplification
b-DNA
-
Automated techniques
SDA, TMA, NASBA TMA-TB, NASBA-HIV, CMV
-
DNA probe on a stationary silicon support
microarray (DNA chip)
-
Factors that can influence exposure to infectious disease
Pathogenicity or virulence of agent, immune states of host, appropriate portal of entry
-
For infectious disease to develop in a host the organism must
penetrate first line of defense, severe natural and adaptive defense
-
Most effective protection of parasitic infections is
antibody dependent cell mediated-cytotoxic
-
Most frequently mediated is
IgG sometimes IgE
-
types of fungal disease
histoplasmosis, aspergillosis, coccidiomycosis
-
Valley fever is the inhalation of dust that contains
arthrospore of coccidioides immitus
-
Define anergy
lack of immunity to antigen
-
histoplasmosis specimen
sputum, blood, tissue
-
Coccidiomycosis specimens
blood and CSF
-
Cryptococcus
Serum and CSF
-
Complement fixation (CF) is the most specific
antibody detection
-
Plays major role in body defenses against viral infections
Interferons
-
Most virulent viruses to humans are
zoonoses
-
Human herpes viruses are
CMV, Epstein-Barr virus, herpes simplex, varicella-zoster, human herpes virus-6
-
Human herpes viruses are large
enveloped DNA virus, replicates in nucleus
-
Varicella-zoster is the same virus that
causes two different types of clinical diseases
-
Primary phase of Varicella-zoster and latent phase
Chicken pox, shingles
-
Complications of VZV
pneumonitis, nephritis, hepatitis, myocarditis, arthritis, and Reye's syndrome
-
Zoster eruptions follow
dermatomes
-
Human herpesvirus 6 targets
T-cells
-
Which antibody is produced in signficant quantities during first exposure to infectious agent
IgM
-
Procedures that specifically evaluate the presence of IgM and IgG used to detect
TORCH taxoplasmosis, other, rubella, CMV, herpes
-
Most common congenital virus?
CMV.
-
Viruses are defended by
interferon
-
Bacteria defended by
Lysozymes and phagocytosis
-
Parasites are defended by
complement, cytotoxic, cellular defense
-
Histoplasmosis is caused by
fungus
-
Aspergillosis is caused by
an opportunistic organism
-
What is the concept of vaccination
deliberating introducing a potentially harmful microbe
-
The purpose of vaccine is to stimulate
active immunity and create immune memory
-
Most vaccines can be divided into two types
Live, attenuated vaccines, non-replacating vaccines.
-
-
vaccines must me specific requirements
produced protective immunity, produce strong and measurable, stable during shelf
-
Dendritic cells can sense pathogens through
pathogen recognition receptors (TLR)
-
Biological warfare pathogen
Smallpox, anthrax, plague, botulism, tularemia, Q fever, Brucellosis,
-
The institute of medicine has ranked which virus vaccine most top priority
CMV
-
Which organism monitors influenza virus
WHO
-
Which HPV strain causes cervical cancer
HPV-16
-
Genetric mutation arise which are called
antigenic drift
-
FluLaval is a
flu vaccine
-
Small pox vaccine are made from live virus called
vaccinia
-
AIDS, CMV have a vaccine
no
-
hay-fever, HPV have vaccines yes or no?
yes
-
Center for biologics Evaluation and research CBER regulate
vaccines
-
Lancefield group A beta hemolysis
S. pyogenes
-
S.pyogenes causitive agent in
Pharngitis, scarlet fever, skin infections, impetigo, necrotizing fascitis.
-
S. pyogenes infections in children, adults
otitis media, sinusitis.
-
High grade streptococcal infection would be
TSS-toxic shock syndrome
-
S.pyogenes what type of bacteria
gram positive cocci
-
S.pyogenes to adhere to cell wall is
fimbriae with lipoteichoic acid
-
M protein which is found in the _____ inhibit ____
fimbriae, phagocytosis and antibody synthesis
-
The virulence factor is caused by
M protein
-
Most frequently used serologic indicator for streptococcal infections
SLO (oxygen labile enzyme
-
Streptolysin S is oxygen stable or lable enzyme and responsible for the alpha or beta hemolysis
stable and beta
-
S. pyogenes typically caused by __ but can be seen in
respiratory droplets, food-borne/milk-borne
-
Carriers have positive cultures but
without serologic evidence of infection
-
Major complication of S. pyogenes
Rheumatic fever
-
Epidemiology of S.pyogenes is
normal flora in the respiratory tract, opportunistic pathogen, rheumatic fever.
-
S. pyogenes (pharyngitis) is
strep throat
-
Most infected patient's S. pyogenes demonsrated increased concentrations of antibody to
SLO
-
S. pyogenes titers rise after about
7 days after exposure, max after 4-6weeks but can be present for up to a year
-
diagnostic evaluation of Streptococcal infections
Throat cultures, rapid strep, ASO (antistreptolysin O, AntiDnas
-
STSS can be caused by
NSAIDS, tampons
-
STSS is caused by
exotoxins
-
Treatment of STSS
penicillin, beta-lactam antibiotics
-
S. agalactiae is
group B, found in blood
-
Most reliable test for recent s.pyogenes skin infection
anti-DNAse B
-
The classic test that demonstrate the presence of streptococcal infections
ASO and anti DNAse B
-
Sphirochetes do not appear to survive in units of
citrated blood at 4C more than 72 hours
-
Spirochete lives in
GI and genital tracts
-
Spirochetes can be viewed using
Darkfield microscopy
-
primary syphilis
painless, lesion (canchre) 1-5 weeks even without treatment) incubation period (last 3 weeks)
-
secondary syphylis
adenopathy (75%), skin lesions (80%), some both primary and secondary overlap 2-8 weeks
-
resolution of secondary syphylis in
2-6 weeks even without therapy
-
late phase (teritary) seen how many years after primary
3-10 years
-
Hutchinsons teeth associated with
congenital syphylis late stage >2 yrs
-
Untreated syphylis late stage involves CNS in about
8% of cases
-
2 classes of treponema antigens
1. specific antibodies 2. nonspecific antibodies
-
Specific antibodies
antigens restricted to 1-2 species of spirochetes
-
nonspecific antibodies
antigens shared by many different spirochetes
-
specific antitreponemal in early and untreated early latent are predominantly
IgM
-
Greatest concentration of IgG seen in
secondary syphylis
-
Non-treponemal antibodies are
Reagin antibodies
-
diagnosis non-treponemal screening
RPR
-
diagnosis treponemal
FTA-ABS,MHA-TP
-
presence of gummas
late (tertiary)
-
diagnostic blood test for syphilis is
Wasserman
-
Are pathogenic treponemes cultivable
no
-
primary incubation period for syphilis
3 weeks
-
syphilis initially treated with
heavy metals
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