PCR
The polymerase chain reaction (PCR) method targets a specific gene for amplification. We currently utilize both conventional and real-time PCR (+/- reverse transcriptase) assays to detect the presence of genes specific to B. pertussis, West Nile Virus, St. Louis Encephalitis (SLE), Eastern Equine Encephalitis (EEE), Western Equine Encephalitis (WEE), Norovirus, and Shiga Toxin-producing E. coli.
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Serology
We use conventional serology techniques to identify serovars/strains of Salmonella and Shigella. The procedure begins by growing specimens on different types of media to give biochemical information. Next, we add various antisera to bacteria isolates and look for agglutination or "clumping," which is indicative of a positive result. Because different Shigella and Salmonella strains have slight variations of antigens on the outside of their outer membranes (e.g. lipopolysaccharide or LPS), only antisera specific for these molecules will induce agglutination. Therefore, this technique allows us to differentiate between the many strains of Shigella and Salmonella. The information gathered here allows epidemiologists to determine which cases may be connected during an outbreak. |
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PFGE
The pulsed field gel electrophoresis (PFGE) method is used to analyze the size of genomic DNA fragments from organisms subjected to restriction enzymes (i.e. enzymes that cut at specific sites within DNA). Our laboratory utilizes PFGE to analyze the genomic DNA of many bacteria, including E. coli, Salmonella, Shigella, Pseudomonas species, Neisseria meningitides strains, Ralstonia species, Acinetobacter species, Staphylococcus aureus (including MRSA), Enterococcus species, Streptococcus species, Serratia species, Pseudomonas fluorescens, and Listeria species. Organisms are grown on selective media, embedded into agarose, lysed, and subjected to restriction enzyme digestion. These so-called agarose "plugs" are inserted into an agarose gel. The gel is then subjected to an electric field, allowing for separation of the individual DNA fragments based on size. Once the gel is stained, it is imaged and the band patterns compared to our vast database. Information obtained from PFGE is used to help identify the strain of the bacteria isolate and helps epidemiologist in hunting down the source of an outbreak. |
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Aaron Benfield, PhD
Team Lead, Molecular Biology
(512) 458-7735
(888) 963-7111 ext. 7735 toll free
email Aaron
Lab phone: (512) 458-7784
(888) 963-7111 ext. 7784 toll free
Laboratory Services Section
Department of State Health Services, Mail Code 1947
PO Box 149347
Austin, TX 78714-9347
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