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Clostridim Difficile

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Clostridim difficile was first isolated in1935 by George and his colleagues from a stool found in a healthy infant by the name of Hall and O’Toole (Heinlen and Ballard, 2010). Yet in 1978, C.difficile was known to be associated as disease in human’s antibiotic-associated diarrhea (Heinlen and Ballard, 2010). It is 2 types of C. difficile: one that exists in spore form and the other in vegetative form. Depending on the type of form it exist in, allows it to grow in a certain environment. It is also a gram positive rod bacterium. When Clostridim difficile exist in a spore form, it can live in harsh conditions and in common sterilization techniques (Heinlen and Ballard, 2010). When C. difficile exist in spore form, it will be resistant to temperature …show more content…

difficile (Heinlen and Ballard, 2010).
In the United States, Clostridim difficile has cost the healthcare system possible more than 1 billion dollars annually, and in developing countries it is the leading cause of illness that occurred in hospitals; with cases that have C. difficile link to it is estimated to cause over 3600 dollars in health care fees (Heinlen and Ballard, 2010). According to data that was reported by Center for Disease Control and Prevention, C. difficile start from a low of 31 cases out of 100,000 people per yer in 1996 to an increase in 2003 with 61cases out of 100,000 people per year (Heinlen and Ballard, 2010). In the United States alone, it have been estimated that 500,000 cases have occurred per year (Heinlen and Ballard, 2010). C. difficile infections (CDI) have also increased by 25% with an estimate of 15,000- 20,000 people die per year in …show more content…

difficile to treat host from this pathogen. They use C. difficile strains, plasmids and conjugal transfer system to conjugate C. difficile gene. C. difficile strain 630 and VPI 10463was used and they first grew it anaerobically in Brain Heart Infusion media or it can be grown on an OXOID agar with 0.5% yeast extract and 0.1% cysteine, BHIS (2013). They also grew the bacteria in modified HySoy Medium by fermentation of 4-81. They then designed synthetic TcdA and TcdB genes using double allelic substitutions in GT catalytic site residues and used 630 toxin genome sequence for the recombinant genes (Donald, et. al, 2013). After synthesis TcdA and TcdB genes with their full length was subcloned as 8.1 kb and 7.1 kb NdeI-BlII fragments into pMTL84123 vector and E.coli strain Stb12 was the recombinant plasmid used (Donald, et. al, 2013). PCR was done using 5’ and 3’ flanking ends of the sites NotI and NdeI RE. Toxin A and B mutants was further mutated to modify or revert the toxins by site-directed mutagenesis using internal 2.5 kb NdeI-HindIII or 3.3 kb NdeI-EcoNI fragment subclones (Donald, et. al, 2013). Clostridim difficile was then conjugated by conjugal transfer of plasmids from E.coli and centrifugation was done to harvest the bacteria (Donald, et. al, 2013). Transformation that appeared on the media or agar was purified onto a BHIS agar and tested genetically modified toxins (Donald, et. al, 2013).

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