Preparation and identification of the oligonucleotide chip for detecting the virulence-associated genotypes and drug resistance of Helicobacter pylori / 中国医学科学院学报

<p><b>OBJECTIVE</b>To develop an oligonucleotide microarray-based method for the simultaneous detection of Helicobacter pylori (Hp) infection, virulence-associated genotypes, and drug resistance.</p><p><b>METHODS</b>Hp was classified into cagA + and cagA- genotypes based on its virulence. Clarithromycin-resistance of Hp was identified by existence of point mutations in 23S rRNA. We constructed an oligonucleotide microarray chip to simultaneously diagnose Hp infection and detect its virulence-associated genotypes and mutations associated with clarithromycin-resistance. The diagnostic accuracy of the constructed microarray was tested with templates of wild type and mutated type.</p><p><b>RESULTS</b>The oligonucleotide chip accurately detected cagA + and cagA- genotypes of Hp, as well as four common point mutations in 23S rRNA related to clarithromycin-resistance.</p><p><b>CONCLUSION</b>Oligonucleotide microarray chip can be used to diagnose Hp infection and test its virulence-associated genotypes and drug resistance simultaneously.</p>

Detection of P53 and K-ras gene mutations in lung cancer with oligonucleotide chip / 生物工程学报

Different factors including hybridization solution components, hybridization temperature, and the concentration and proportion of the labelled primer, which affected the sensitivity and specificity of single mutation identification, were exploited. Asymmetric PCR increased the hybridization sensitivity, and the asymmetric multi-PCR did not affect the specificity, while the sensitivity was improved a little. Among 30 lung cancer samples detected with the oligonucleotide microarray, 12 was found P53 gene mutations and 5 had K-ras gene mutations. The P53 gene mutations identified by the oligonucleotide microarray was proved 80% same as the sequencing results. The obvious statistical relations of K-ras and P53 gene mutations with tumor type, tumor stage and smoking were not obtained because of less samples and mutation sites.