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Lab diagnosis
Lectures 19-20
Question | Answer |
---|---|
non-specific signs/test | physical: fever, pain, swelling, redness general lab tests: CBC (increased WBC), non-specific (increased RBC sedimentation rate, elevated CRP), localized changes (increased protein in CSF, abnormal liver function) |
accuracy vs. precision | precision: how consistent are the values accuracy: how close are the test values to the true value |
predictive value | a measure of the times that the value is the true value |
impact of incidence on sensitivity | a higher prevalence leads to better performance in a test |
immunodiagnosis of acute infections | draw an acute serum (before 14 days), draw a convalescent serum 3-6 weeks later (look for a 4 foldor greater increase due to IgG). Most use ELISA now so 4 fold increase in optical density |
If you miss the acute titer... | draw a convalescent titer at the peak (3-6 weeks after start of infection) A very high convalescent titer=infection presumptively confirmed. A single positive Ab does not allow definitive confirmation (exception: lyme diseae, HIV, hepatitis) |
diagnosis of chronic infections | HIV/lyme disease: western blot Hepatitis B: several Ab markers HIV/Hepatitis B and C: can also check for the presence of the Ag by PCR and Ag immunoassay for Hepatitis B |
Test for congenital infection | check Ab titer against agents suspected, test cord blood serum at birth, test again after 3-4 months. Baby negative: IgM not present, IgG from mom will disappear at 50% drop rate every month Baby positive: IgM positive and IgG will go up |
Third generation immunoassays | sensitivity and specificity are typical of enzyme immunoassays single analyte tests: easily detect one thing, sold phase or membrane based, convenient to perform |
Fourth generation assay | detect two things or use novel technology simultaneous: Ag and Ab or multiple Ag kites made with superior reagents: monoclonal Ab or recombinant Ag New improved detection technology: time resolve fluorescence, immunoprobe for fiber optics |
First molecular nucleic acid amplification method | target amplification: PCR target fingerprint-->amplification and probe capture-->detection |
emerging diagnositc technologies | microarrays: for microbial identification and typing, detection of host responses to infection. Unique to microarrays (Can do 10 to 20000 tests on a simgle specimen) |
types of arrays | microbe oriented-look for pathogen host oriented-check for the patient's response to infection, check for host characteristics that predict worse outcomes or problems with a particular drug |
technology summary | cultures give positive vs negative answers chemistry styles give grey answers: color production with varying degrees of intensity, can vary from test to test important that patients realize the possibility of false pos/neg |
disease prevalence affects predictive values | increasing prevalence increases the PVP/PVN |
situations for rejection of specimens that are beyond salvage | misidentification, culture specimens received in fixate, dried out specimens on swab, insufficient quality |
criteria for rejection of seriously compromised specimens | improper storage temperature, improper transport medium, improper transport time, leaky specimen |
organisms requiring prompt culture if specified by doctor | Haemophilus ducreyi, anaerobes in regular transport tubes, Neisseria gonorrhoeae in joint fluid |
contaminated specimens | expected: contain normal flora that needs to be distinguished from pathogens unexpected: sterile with introduced bacteria |
blood cultures | continuous bacteremia: volume dependent, collect 20-30 ml per set, 2-3 sets per episode intermittent bacteremia: 2-3 sets before antibiotics collect each set from a new venopuncture site: two sticks is a minimum |
things to avoid in blood culture collection | disinfecting collection site with alcohol only(iodophor for 2 minutes),initially collecting >5 blood culture sets to diagnose a single clinical episode,collecting only one bottle per set(1 aerobic and 1 anaerobic bottle),collecting all from 1 needle stick |
blood culture issues | 3% increase in yield/mL (collect 20mL/set and 2-3 sets), central venous catheters (semiquantative culture of CVC tip: >15 significant), postmortem blood rarely useful |
true positives vs. contaminants | true positives: multiple sets and bottles are positive, positives arise within 3 days, organism isolated is typical pathogenContaminant: only one bottle positive, organism normal skin type flora, takes more than 5 days to grow |
CSF | wear mask to avoid normal oral flora in tubes, do gram stain drom cell count tube to protect culture tube (should see organism in gram stain 70% of time) contaminant have negative gram stain and <3 colonies air contaminants are not on streak line |
detecting agents of pneumonia | best sample is detection of lung tissue collected in OR, second best is saliva free sputum collected by bronchoscopy, third best is good cough sputum |
cough sputum cultures | avoid saliva (small quantities of thin clear liquid is from mouth...will have squamous epithelial cells and few WBC) |
urine cultures | bacterial growth: doubles every 30 minutes pathogen and vaginal flora may mingle (refrigerate to preserve true numbers) |
quantitation and significance of bateruria | likely to be significant 80% of time (100000 CFU/ml of one organism) in special circumstances 10^2 or 10^3 CFU/mL may be significant in men or symptomatic women |
urine collection | clean catch, first morning specimen refrigerate or put in special preservation tube |
urine culture preservation | refrigeration alternative, boric acid based, mildly toxic, dilution onto media initiates growth. Colony counts stable for 8-24 hours at 20C |
quantitation and significance of bacteruria | contaminated specimens (high quantities of >3 organisms), correlation with significant bacteriuria: gram stain (at least 2 bacteria per high power field has 90% correlation and significance) |
Unsuitable specimens for lab detection of UTI's | foley catheter tips, urine from catheter bag, unsupervised clean catch specimens from female patients who have not had instructions, urine over 2 hours old at 20C |
community acquired diarrhea | occurs before 3 days in hospital, may be due to parasites, enteric bacterial pathogens, viruses |
Hospital patients with diarrhea | C. difficile, in hospital over 3 days, on broad spectrum antibiotics or chemotherapy |
wound specimens | tell lab the anatomical site, surface wounds have skin flora as pathogens and contamination, don't sample superficial pus, deep wound sample okay |
Do's and don'ts for likely contaminated surface wounds | don't swab superficial layers or put swab through contaminated superficial layer trying to go deeper, don't use surgical techniques to take a superficial specimen with surface contamination |
rejections related to anaerobes | don't culture from these anaerobes: fecal contaminated specimens, midstream and catheter urine, expectorated sputum, throat nose orophayngeal swabs, gastric contents, vaginal and cervial swabs, superficial material from skin |
Minimum inhibitory concentration | least antibiotic concentration that inhibits growth |
problems with MIC tests | can be expensive, each drug takes multiple tubes |
disk diffusion test | mueller hinton agar plate seeded with organism, position paper disks impregnanted with antibody, measure zone in mm |
problems with disk diffusion | overnight incubation required, average accuracy 92%, increasing number of drug/bug specific contraindications limits ability to be a universal test |
E-test | Antibiotic is impregnanted onto a rectangular plastic strip, like disk diffusion, but strip is placed on the agar |
Extended spectrum beta lactamase | plasmid mediating beta lactamases that confer resistance to penicillin, cephalosporins, and aztreonam. Mainly in klebsiella and E.Coli |
ESBL enterobacteriaceae | appear susceptible in vitro to second generation cephalosporins and resistant to first and many of the third generation |
inducible beta lactamases | enzyme conferring resistance to penicillins and cephalosporins in certain gram negative rods. Delayed and develops slowly. May occur with klebsiella, citrobacter, enterobacter, E.coli, aeromonas, pseudomonas, and some others |
Inducible AmpC beta lactamase | just became ready for limited routine testing, on plasmid in some enterobacteriaceae |
Klebsiella Pneumoniae Carbapenemase | beta lactamase, confers resistance to all beta lactams, occur in enterobacteriaceae, most commonly klebsiella. Rarely in pseudomonas |
lab detection of KPC producers | problem: some isolated demonstrate low level carbapenem resistance, some automated systems fail too detect low level resistance |
When to suspect a KPC producer | enterobacteriaceae (esp. Klebsiella) that are resistant to extended spectrum cephalosporins, and resistant or intermediate to cefoxitin and cefepime |
empirical antimicrobial therapy is based on | historical susceptibility patterns, body site and acuity of disease, likely organis, involved, clinical trial data |