Biomedical subjects

Erik Henchal

Publications and source records attributed to Erik Henchal.

3 recordsLinked to original sources

PubMed2003

Use of denaturing high-performance liquid chromatography to identify Bacillus anthracis by analysis of the 16S-23S rRNA interspacer region and gyrA gene.

Denaturing high-performance liquid chromatography (DHPLC) was evaluated as a method for identifying Bacillus anthracis by analyzing two chromosomal targets, the 16S-23S intergenic spacer region (ISR) and the gyrA gene. The 16S-23S ISR was analyzed by this method with 42 strains of B. anthracis, 36 strains of Bacillus cereus, and 12 strains of Bacillus thuringiensis; the gyrA gene was analyzed by this method with 33 strains of B. anthracis, 27 strains of B. cereus, and 9 strains of B. thuringiensis. Two blind panels of 45 samples each were analyzed to evaluate the potential diagnostic capability of this method. Our results show that DHPLC is an efficient method for the identification of B. anthracis.

Bacillus anthracis↗

PubMed2003

Detection and identification of ciprofloxacin-resistant Yersinia pestis by denaturing high-performance liquid chromatography.

Denaturing high-performance liquid chromatography (DHPLC) has been used extensively to detect genetic variation. We used this method to detect and identify Yersinia pestis KIM5 ciprofloxacin-resistant isolates by analyzing the quinolone resistance-determining region (QRDR) of the gyrase A gene. Sequencing of the Y. pestis KIM5 strain gyrA QRDR from 55 ciprofloxacin-resistant isolates revealed five mutation types. We analyzed the gyrA QRDR by DHPLC to assess its ability to detect point mutations and to determine whether DHPLC peak profile analysis could be used as a molecular fingerprint. In addition to the five mutation types found in our ciprofloxacin-resistant isolates, several mutations in the QRDR were generated by site-directed mutagenesis and analyzed to further evaluate this method for the ability to detect QRDR mutations. Furthermore, a blind panel of 42 samples was analyzed by screening for two mutant types to evaluate the potential diagnostic value of this method. Our results showed that DHPLC is an efficient method for detecting mutations in genes that confer antibiotic resistance.

Anti-Infective Agents↗

PubMed2002

Denaturing HPLC for identifying bacteria.

Denaturing HPLC (DHPLC) is used in a wide variety of genetic applications. Here we introduce a new application for this technique, the identification of bacteria. We combined the capability of DHPLC to detect sequence variation with the principles of rRNA genotyping analysis to develop a high-throughput method of identifying microorganisms. Thirty-nine bacterial species from a broad spectrum of genera were tested to determine if DHPLC could be usedfor identification. Most (36 of 39) species of bacteria had a unique peak profile that could be used as a molecular fingerprint. Furthermore, a blind panel of 65 different bacterial isolates was analyzed to demonstrate the diagnostic capability of this method to specifically identify Yersinia pestis and Bacillus anthracis. All the Y. pestis samples (10 of 10) and the majority of B. anthracis samples (12 of 14) were correctly identified. The procedure had an overall specificity of 100%, overall sensitivity of 91.7%, and a predictive value of 96.9%. The data suggest that DHPLC of products spanning regions of genetic variability will be a useful application for bacterial identification.

Bacteria↗