摘要
应用巢式聚合酶链反应技术，通过两种引物（共四对）分别对伤寒沙门氏菌鞭毛Ｈ和Ｖｉ抗原基因扩增，并用非伤寒沙门氏菌作对照。实验表明，两种基因的扩增产物均是特异的，Ｈ抗原基因引物仅对伤寒沙门氏菌鞭毛抗原基因扩增，Ｖｉ抗原基因引物对伤寒沙门氏菌和丙型副伤寒沙门氏菌Ｖｉ抗原基因扩增，余均为阴性。扩增Ｖｉ抗原基因比扩增Ｈ抗原基因敏感，当反应体系中达０．５个菌细胞时，Ｖｉ抗原基因就可检出，而鞭毛抗原基因则需５个菌细胞。检测９２例伤寒患者血液标本，血培养阳性２８例（３０．４３％），巢式ＰＣＲ检测Ｖｉ抗原基因阳性３３例（３５．８６％），Ｈ抗原基因阳性３１例（３３．７０％）。伤寒发病早期，当机体伤寒特异性抗体尚处于低水平时，应用巢式ＰＣＲ检测Ｖｉ抗原基因，可提高伤寒的诊断率。
A nested PCR assay for the detection of Salmonella typhi was developed by using four sets of primers for amplification of the flagellin gene and Vi antigen gene.Our results showed that only S.typhi strains were positive by PCR with primers based on the flagellin gene(herein as refer to HDNA)and S.typhistrains along with a Salmonella paratyphi C strain were positive by PCR with primers based on the Vi antigen gene(Vi-DNA).The sensitivity of Vi-DNA by PCR was higher than that of H-DNA by PCR,the former was approximately 0.5 microorganism and the latter was 5 microorganisms.Among 92 samples from patients with typhoid fever,28(30.43%)showed positive for Salmonella typhi by blood culture,33(35.86%)Vi-DNA PCR assay,31(33.70%)by H-DNA PCR assay.At the early phase of typhoid fever,nested PCR for the detction of Vi antigen gene was capable to increase the positive value for the early diganosis of typhoid fever.
出处
《中国卫生检验杂志》
CAS
1998年第4期201-205,共5页
Chinese Journal of Health Laboratory Technology
基金
江苏省卫生厅资助
