Medical Mircobiology: PPT 1

top 3 ranked world infectious diseases:

Diabetes mellitus, Tuberculosis, HIV/AIDS

TDR

totally drug resistant

MDR

multidrug resistant

antibiotics

do not work on viral infections

post antibiotic era

bacteria have become so resistant to current treatments that common illnesses cannot be effectively treated

prevention

requires no drugs

requires no hospital

works on the unknown

3 pillars of prevention

awareness, implementation, and consistency

ALARA

as low as reasonably achievable

neglected epidemic

herpes epidemic

public prevention bottom lines

plain to understand, easy to do, short to listen to, make sense to believe it, desire to keep doing it

personal prevention

be clean, be moderate, be separate, diverse diet

hospital prevention

Separate, Minimization, Identification, Let Extended rest occur

environmental prevention

be separate, be clean, be moderate, allow for diversity in crops

visual diagnosis

using visual tools to aid in the identification and diagnosis of pathogens (fever, rash, damaged tissue, behavioral changes, pain, etc.)

microscopy

the use of microscopes to identify pathogens in patient sample

negative stain

uses an acidic dye

capsule stain

combination of a negative stain followed by a simple stain

gram stain

crystal violet (1min)

iodine (1min)

alcohol(30sec)

safranin (1min)

acid fast stain

carbocalfushin (1 min)

steam (5 min)

acid-alcohol (1min)

methylene blue (1 min)

acid fast bacteria

Mycobacteria and Nocardia

endospore stain

malachite green (1 min)

steam (5 min)

water (1 min)

saphranin (1 min)

rapid strip test

tests bacteria’s nutrional requirements within one test

sensitive, intermediate, resistant

likelihood of treatment to work on a patient

MIC

minimum inhibitory concentration

zone of inhibition

area around disc that bacteria cannot grow near antimicrobial or antibiotics

cytopathic effect

viral host cell damage

cytopathic effect occurs in 3 ways:

viral overload, cytocidal effects, and noncytocidal effects

polyclonal antibody

inject antigen into the host, stimulate immune response, purify antibody from serum

antibodies can recognize multiple epitopes

monoclonal anitbody

animal produces immune response with B cells, pruify B cells that produce desired antibody

anitbody recognizes one epitope

primary antibody

binds to desired antigen

secondary antibody

binds to another specific kind of antibody

usually tagged to visualize easily

fluorescent antibodies

label antibody; can visualize anything the antibody binds to

serotyping

slide agglutination test; use of antibodies to break organisms into groups

flow cytometry

moving cell measurement; cells sent down a tube one by one to look for specific properties

must be in liquid suspension

ELISA

Enzyme Linked ImmunoSorbent Assay

enzyme added to attach to antibody, if antibody attaches to antigen it can be seen by adding serum

antigen capture ELISA

anitbodies used to capture specific antigen; detecting antigen

antibody capture ELISA

antigen is used to capture specific antibodies; detecting immune repsonses

western blot

look for specific proteins in specific organisms

IgM

immune repsonse in first 30 days of infection

IgG

past immune response; after intial repsonse

Direct DNA detection

gel electrophoresis, southern, northern, microarray

Indirect DNA detection

PCR, RT-PCR, Real Time PCR

identification of DNA

fingerprinting, RFLP, sequencing, microarray

probes

looking for fragments of DNA that can be used to look for specific sequences

in situ hybridization

probe tissue for DNA/RNA

southern

probe for DNA

northern

probe for RNA

microarray

probe for hundreds of fragments, sample labeled, probe is bound

PCR

amplify DNA up into DNA

RT-PCR

amplifies RNA to DNA

Real Time PCR

combination of southern with PCR; can determine moment PCR became positive (Q-PCR)

PCR steps

denaturation, annealing of the primer, and amplification

nested PCR

sequential PCR using two sets of primers, more sensitive than regular PCR, less false positives

semi-nested PCR

only one primer changes so more false positives than nested by still less than PCR

multiplex PCR

doing more than one PCR at the same time in the same tube

fingerprinting

short primers used to look for specific DNA sequences; DNA

RFLP

restriction fragment length polymorphism

compare banding patterns to look for matches

PCR followed by RFLP reduce number of false postives

PFGE

pulse field gel electrophoresis

used to visualoze large DNA fragments

MALDI-TOF-MS

use laser to release protein fragments, use mass spectrometer to measure properties of ions

can match genus, species, and sometimes strain