Fundamentals Flashcards
Why utilize a diagnostic laboratory?
- to determine if an infectious agent is present (does my patient/herd/flock have an infection?)
- to obtain an etiological diagnosis (what organism am I dealing with?)
- to guide antimicrobial therapy (what is the most appropriate drug to use?)
- THE ONUS IS ON YOU TO UNDERSTAND, & THEREFORE BE ABLE TO EXPLAIN TO YOUR CLIENT, WHY A LABORATORY DIAGNOSTIC TEST IS IMPORTANT!
What are other important functions of diagnostic labs?
- disease surveillance (passive): antimicrobial resistance; trend/outbreak ID
- regulatory: testing for reportable diseases; export/import requirements; public health/food safety
- research: diagnostic submissions are a valuable source of research materials
What is a colony?
blue circles are colonies
- colonies on an agar plate are POPULATIONS of bacteria
- clonal population which descended from a single viable organism (the CFU)
What is a CFU?
colony forming unit: when one cell expands to form entire colony (the ancestor organism)
Assuming a 20 minute generation time, if we start with 1 CFU, how many organisms will we have after 1 day of unimpeded division?
1->2->4->8->16->32->64->128->256->512->1024->2048-> 4096 -> etc.
4.72x10^21
this is just a theoretical b/c would run out of nutrients, etc.
What does a colony theoretically represent & what can it be used to estimate?
- presence of a single viable progenitor organism & is therefore a clonal population
- Can therefore be used to estimate the number of bacteria (CFU) in a sample by making dilutions and plating them until you can count organisms per plate
Why do we care how many organisms are present on a plate?
- establishing clinical significance (from free catch urine, UTI is defined as >50,000 CFU/ml)
- to identify contaminants (identifying dominant organisms in mixed cultures - probably the dominant organism is the one causing the problem)
- to standardize laboratory tests (some assays require testing a specific number of organisms)
How do you streak out plates for isolation?
- 4 streak method
- objective is to get isolated colonies
How is the semi-quantitative method ranked on plates?
- scant = < 10 colonies on 1st streak
- 1+ = > 10 colonies on 1st
- 2+ = > 10 colonies on 2nd
- 3+ = > 10 colonies on 3rd
- 4+ = > 10 colonies on 4th
What is an isolate?
- a pure culture (clonal)
- derived from a single colony
- genetically homogenous
- suitable for additional characterization: identification, susceptibility testing
How are culture media selected?
- selection of a primary media depends on what organisms you are interested in isolating - what is your target pathogen (blood agar & MacConkey are pretty standard for routine cultures)
- introduction of selective & differential media facilitates a presumptive identification on primary culture
What is selective media used for?
- to preferentially isolate particular taxa (contains chemicals to inhibit the growth of non-target organisms)
What does CNA media contain?
Colistin & Nalidixic acid
What does MacConkey media contain?
Bile salts & crystal violet
What does Campy-BAP media contain?
Bacitracin, novobiocin, colistin, cephalothin, polymyxin B
What does CNA media select for?
gram - positives
What does MacConkey media select for?
gram - negative enterics (ex: E. coli)
What does Campy- BAP media select for?
Campylobacter jejuni
What does CNA media select against?
gram-negatives
What does MacConkey media select against?
gram positives
What does Campy-BAP media select against?
most other bacteria (other than Campylobacter jejuni)
What is differential media used for?
- exploits physiological properties of organisms of interest to produce unique colony morphologies (often colourimetric)
What is the differential ingredient in MacConkey media?
lactose
What is the differential ingredient in XLD media?
Ferric ammonium citrate