以專一性引子進行創傷弧菌的鑑別

Specific Primers for Rapid and Sensitive Identification of Vibrio Vulnificus

由創傷弧菌的藍色螢光蛋白基因序列設計一組引子，以聚合酵素鏈反應方法，這組引子可從所有來自臨床的創傷弧菌擴增獲得預期的核酸片段251鹼基對，而不會由其他的臨床致病菌中產生任何核酸片段。這組引子可偵測創傷弧菌染色體含量少至1豪微微克或少至60個細菌數。在生物體樣品的菌體檢測評估中，以血清和傷口抽取液共22件進行聚合酵素鏈反應檢測。在14件細菌培養有細菌存在的樣品中，有13件在聚合酵素鏈反應檢測中呈現正反應，比率高達92.9%。然而，在8件細菌培養無細菌存在的樣品中，有3件在聚合酵素鏈反應檢測可呈現正反應，比率有37.5%之高。在進行細菌鑑別時，這組引子除了具有高專一性的性質外，並且在一階段式的聚合酵素鏈反應仍具有高靈敏性之特質；因此，這組引子是一對可用來進行鑑別創傷弧菌的重要引子。

A primer pair designed from vvbfp （Vibrio vulnificus blue fluorescent protein） of V. vulnificus amplified a 251 bp PCR fragment from all clinical isolates of V. vulnificus and failed to amplify any fragment from other clinical pathogens. The sensitivity of the primer pair is as little as 1 fg of chromosomal DNA and 60 cells of V. vulnificus. In a scientific evaluation of specimens, a total of 22 samples including serum specimens and wound aspirates from septicemic mice were screened by PCR. Thirteen （92.9%） of the 14 V. vulnificus culture-positive specimens gave positive results. Three （37.5%） of the 8 culturenegative specimens yielded positive results. The primer set gives high sensitivity and specificity in a one-step PCR protocol, an important criterion for unambiguous bacterial identification.