广东海洋大学学报
廣東海洋大學學報
엄동해양대학학보
JOURNAL OF GUANGDONG OCEAN UNIVERSITY
2013年
3期
52-55
,共4页
陈福艳%程光平%欧阳贤华%陈晓汉%梁万文%韦友传%黄婷%杨学明%陈明%王瑞%李丽萍%韦明利
陳福豔%程光平%歐暘賢華%陳曉漢%樑萬文%韋友傳%黃婷%楊學明%陳明%王瑞%李麗萍%韋明利
진복염%정광평%구양현화%진효한%량만문%위우전%황정%양학명%진명%왕서%리려평%위명리
双重PCR%创伤弧菌%哈氏弧菌
雙重PCR%創傷弧菌%哈氏弧菌
쌍중PCR%창상호균%합씨호균
duplex PCR%Vibrio vulnificus%Vibrio harveyi
根据创伤弧菌（Vibrio vulnificus）的溶细胞毒素基因序列和哈氏弧菌（Vibrio harveyi）的toxR基因序列，分别设计并合成两对特异性引物，通过PCR反应条件优化，测试两种菌的特异性和敏感性，建立双重PCR方法，同时快速检测V. vulnificus和V. harveyi。结果表明：纯培养V. vulnificus和V. harveyi的检测灵敏度分别是12 cfu/mL和18 cfu/mL，与无乳链球菌、海豚链球菌、副溶血弧菌及美人发光杆菌无交叉反应；此PCR 检测方法具有良好的特异性、敏感性，具快速、高效等优点，对细菌V. vulnificus和V. harveyi诊断与防治具有较好的临床应用性。
根據創傷弧菌（Vibrio vulnificus）的溶細胞毒素基因序列和哈氏弧菌（Vibrio harveyi）的toxR基因序列，分彆設計併閤成兩對特異性引物，通過PCR反應條件優化，測試兩種菌的特異性和敏感性，建立雙重PCR方法，同時快速檢測V. vulnificus和V. harveyi。結果錶明：純培養V. vulnificus和V. harveyi的檢測靈敏度分彆是12 cfu/mL和18 cfu/mL，與無乳鏈毬菌、海豚鏈毬菌、副溶血弧菌及美人髮光桿菌無交扠反應；此PCR 檢測方法具有良好的特異性、敏感性，具快速、高效等優點，對細菌V. vulnificus和V. harveyi診斷與防治具有較好的臨床應用性。
근거창상호균（Vibrio vulnificus）적용세포독소기인서렬화합씨호균（Vibrio harveyi）적toxR기인서렬，분별설계병합성량대특이성인물，통과PCR반응조건우화，측시량충균적특이성화민감성，건립쌍중PCR방법，동시쾌속검측V. vulnificus화V. harveyi。결과표명：순배양V. vulnificus화V. harveyi적검측령민도분별시12 cfu/mL화18 cfu/mL，여무유련구균、해돈련구균、부용혈호균급미인발광간균무교차반응；차PCR 검측방법구유량호적특이성、민감성，구쾌속、고효등우점，대세균V. vulnificus화V. harveyi진단여방치구유교호적림상응용성。
As Vibrio vulnificus and Vibrio harveyi mainly affects marine cultured animals, it is necessary to establish a rapid detection method. According to the cytolysin gene sequence of V. vulnificus and toxR gene sequence of V. harveyi, two pairs of primers were designed and synthesized. The diagnosis of duplex polymerase chain reaction (PCR) for rapid identification of V. vulnificus and V. harveyi was established by optimization of PCR conditions and by testing specificity and sensitiveness. The duplex PCR test results showed that two specific and amplified fragments were produced which sizes were 128 bp for V. vulnificus and 211 bp for V. harveyi, and no cross-reaction tests were done with Streptococcus agalactiae, Streptococcus iniae, Bibrio parahemolyticus, and Vibrio damsela. The lowest concentrations of 12 cfu/mL for V. vulnificus and 18 cfu /mL for V. harveyi could be detected. The duplex PCR would be a specific and sensitive method which was timesaving, laborsaving, quick and high-efficient for identification of duplex V. vulnificus and V. harveyi.
