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一种用于检测血液样本中伤寒沙门氏菌等沙门氏菌血清型聚合酶链反应产物的高灵敏度快速非放射性方法。

A highly sensitive and fast non-radioactive method for the detection of polymerase chain reaction products from Salmonella serovars, such as Salmonella typhi, in blood specimens.

作者信息

Cocolin L, Manzano M, Astori G, Botta G A, Cantoni C, Comi G

机构信息

Dipartimento di Scienze degli Alimenti, Facoltà di Agraria, Università degli Studi di Udine, Italy.

出版信息

FEMS Immunol Med Microbiol. 1998 Nov;22(3):233-9. doi: 10.1111/j.1574-695X.1998.tb01211.x.

Abstract

A polymerase chain reaction based test was developed for the detection of Salmonella spp. in blood specimens. After amplification of a 389 bp-polymerase chain reaction product from the invA gene, a microtiter plate hybridization assay was performed. The protocol described allowed the detection of six to seven copies of the Salmonella typhi genome, as determined by serial dilutions of DNA from S. typhi. Eighteen blood specimens from artificially infected rats and 22 blood specimens from patients were analyzed to validate the method. Considering that the most frequent Salmonella serovar isolated from blood in case of bacteremia is S. typhi, the polymerase chain reaction-microtiter plate hybridization technique could be used as a novel, rapid diagnostic method for typhoid fever, particularly when standard culture assays are negative.

摘要

开发了一种基于聚合酶链反应的检测方法,用于检测血液标本中的沙门氏菌属。从invA基因扩增出389 bp的聚合酶链反应产物后,进行了微量滴定板杂交试验。所述方案能够检测到伤寒沙门氏菌基因组的六至七个拷贝,这是通过对伤寒沙门氏菌DNA进行系列稀释确定的。分析了18份人工感染大鼠的血液标本和22份患者的血液标本以验证该方法。鉴于在菌血症情况下从血液中分离出的最常见沙门氏菌血清型是伤寒沙门氏菌,聚合酶链反应-微量滴定板杂交技术可作为一种新型、快速的伤寒热诊断方法,特别是在标准培养试验为阴性时。

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