Finals: Midterm 1 Content Flashcards
pathogen
- a microbe that is capable of causing host damage
pathogenicity
- the ability to produce disease
virulence (2)
- degree of pathogenicity
- only expressed in a susceptible host
gram negative bacteria (2)
- thin peptidoglycan cell wall, surrounded by outer membrane containing lipopolysaccharide
- double membrane
gram positive bacteria (2)
- lack an outer membrane
- surrounded by thick peptidoglycan cell wall
Koch’s Postulate: First Postulate
- the microorganism must be found in abundance in all organisms suffering from the disease, but should not be found in healthy organisms
Koch’s Postulates: Second Postulate
- the microorganism must be isolated from a diseased organism and grown in pure culture
Koch’s Postulate: Third Postulate
- the cultured microorganism should cause disease when introduced into a healthy host
Koch’s Postulates: Fourth Postulate
- microorganism must be re-isolated from the inoculated diseased experimental host and identified as being identical to the specific causative agent
Challenges to Koch’s Postulates: First (3)
- host susceptibility varies in human and animal populations due to a variety of factors
- opportunistic bacteria can be found in healthy hosts too
- may be difficult to isolate bacteria from certain areas, such as brain and bones
Challenges to Koch’s Postulates: Second Postulate (2)
- not all bacteria can be cultured in pure form
- many bacteria undergo genetic and phenotypic changes in culture environments
Challenges to Koch’s Postulates: Third Postulate (2)
- not all bacterial species are equally virulent
- closely related bacterial strains can be very different
Challenges to Koch’s Postulates: Fourth Postulate
- re-inoculation of a host may not be possible or ethical
Modern Approaches to Satisfy Koch’s Postulates (4)
- PCR
- immunohistochemistry and immunofluorescence
- ELISA
- eliminate the pathogen and prevent/cure the disease
Modern Approaches to Satisfy Koch’s Postulates: PCR
- PCR and 16S sequencing can be used to amplify and detect bacteria during culturing
Modern Approaches to Satisfy Koch’s Postulates: PCR/16S Sequencing steps (2)
- 16S RNA primers are designed to detect conserved regions, with unique regions in-between that act as barcodes to identify the bacteria
- PCR is done to amplify the bacterial nucleic acid for detection
Modern Approaches to Satisfy Koch’s Postulates: Immunohistochemistry (2)
- microscopic images of immunohistochemical staining can be used to visualize the pathogen in the tissue directly
- uses of antibodies against the bacteria with attached reporter enzymes
Modern Approaches to Satisfy Koch’s Postulates: Immunofluorescence (2)
- micrographs of cells with fluorescently labelled antibodies against cell markers and bacterial
- cells infected with bacteria will exhibit colocalization of fluorescent markers, while healthy cells will not
Modern Approaches to Satisfy Koch’s Postulates: ELISA (2)
- use of fluorescently labelled antibodies against bacteria to detect pathogens in infection site without culturing
- Western Blot/SDS-PAGE can be used subsequently
ELISA methods (3)
- direct ELISA
- indirect ELISA
- capture/sandwich ELISA
Modern Approaches to Satisfy Koch’s Postulates: Eliminate the Pathogen and Prevent/Cure the Disease; Why didn’t Koch try this?
- antibiotics did not exist at the time, so treatment was a lot more complicated and time-consuming
virulence factors
- bacterial product or strategy that contributes to the ability of the bacterium to survive in the host/cause infection
Molecular Version of Koch’s Postulates: First Postulate
- gene for virulence should be present in the strain of bacteria that cause disease and absent in avirulent strains
Molecular Version of Koch’s Postulates: Second Postulate
- (i) knocking out or disruption the gene should reduce virulence, and (ii) introduction of the cloned gene into an avirulent strain should render the avirulent strain virulent
Molecular Version of Koch’s Postulates: Third Postulate
- expression of the gene should be demonstrated in human or a relevant model