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通过聚合酶链反应快速鉴定A型和B型葡萄球菌剥脱毒素基因

Rapid identification by polymerase chain reaction of staphylococcal exfoliative toxin serotype A and B genes.

作者信息

Sakurai S, Suzuki H, Machida K

机构信息

Division of Molecular Genetics, Jikei University School of Medicine, Tokyo, Japan.

出版信息

Microbiol Immunol. 1995;39(6):379-86. doi: 10.1111/j.1348-0421.1995.tb02216.x.

Abstract

A new system was designed to detect staphylococcal exfoliative toxin A (ETA) and B (ETB) genes by the polymerase chain reaction (PCR). The primer pairs for the ETA gene (eta) were 20 and 20-mer, and its PCR product was a 741-bp eta fragment, while the primer pairs for the ETB gene (etb) were also 20 and 20-mer, and its PCR product was a 629-bp etb fragment. When these primers were simultaneously used in the PCR, the two types of ET were clearly detected as two bands in an ETA and ETB double-producer using only one colony within 3 hr. We examined 66 strains of Staphylococcus aureus isolated from patients with staphylococcal scalded skin syndrome (SSSS) and compared the results obtained by ELISA and PCR. The same results were obtained for 56 of the strains, i.e., 30 strains were ETA producers, 20 strains were ETB producers, and 6 strains were double-producers. However, positive results were obtained for 5 of the 10 non-ET-producing strains. Two of these strains were judged by PCR as ETA producers and three as ETB producers. Thus, PCR is very sensitive and rapid in detecting ETA and ETB gene fragments in colonies isolated from patients with SSSS.

摘要

设计了一种新系统,通过聚合酶链反应(PCR)检测葡萄球菌剥脱毒素A(ETA)和B(ETB)基因。ETA基因(eta)的引物对为20聚体和20聚体,其PCR产物是一个741bp的eta片段,而ETB基因(etb)的引物对也是20聚体和20聚体,其PCR产物是一个629bp的etb片段。当这些引物同时用于PCR时,在仅使用一个菌落的情况下,3小时内就能在ETA和ETB双产生菌中清晰地将两种类型的ET检测为两条带。我们检测了从葡萄球菌烫伤样皮肤综合征(SSSS)患者中分离出的66株金黄色葡萄球菌,并比较了ELISA和PCR的检测结果。56株菌株得到了相同的结果,即30株为ETA产生菌,20株为ETB产生菌,6株为双产生菌。然而,10株非ET产生菌中有5株得到了阳性结果。其中2株经PCR判定为ETA产生菌,3株为ETB产生菌。因此,PCR在检测从SSSS患者分离出的菌落中的ETA和ETB基因片段时非常灵敏且快速。

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