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运用免疫磁珠分离法和聚合酶链式反应/DNA探针技术从牛粪便中检测大肠杆菌O26、O103、O111和O145血清型。

Detection of Escherichia coli serogroups O26, O103, O111 and O145 from bovine faeces using immunomagnetic separation and PCR/DNA probe techniques.

作者信息

Jenkins C, Pearce M C, Smith A W, Knight H I, Shaw D J, Cheasty T, Foster G, Gunn G J, Dougan G, Smith H R, Frankel G

机构信息

Laboratory of Enteric Pathogens, Health Protection Agency, 61 Colindale Avenue, London, NW9 5HT, UK.

出版信息

Lett Appl Microbiol. 2003;37(3):207-12. doi: 10.1046/j.1472-765x.2003.01379.x.

Abstract

AIMS

The aim of this study was to isolate Escherichia coli O26, O103, O111 and O145 from 745 samples of bovine faeces using (i) immunomagnetic separation (IMS) beads coated with antibodies to lipopolysaccharide, and slide agglutination (SA) tests and (ii) PCR and DNA probes for the detection of the Verocytotoxin (VT) genes.

METHODS AND RESULTS

IMS-SA tests detected 132 isolates of presumptive E. coli O26, 112 (85%) were confirmed as serogroup O26 and 102 had the VT genes. One hundred and twenty-two strains of presumptive E. coli O103 were isolated by IMS-SA, 45 (37%) were confirmed as serogroup O103 but only one of these strains was identified as Verocytotoxin-producing E. coli (VTEC). Using the PCR/DNA probe method, 40 strains of VTEC O26 and three strains of VTEC O103 were isolated. IMS-SA identified 21 strains of presumptive E. coli O145, of which only four (19%) were confirmed as serogroup O145. VTEC of this serogroup was not detected by either IMS-SA or PCR/DNA probes. E. coli O111 was not isolated by either method.

CONCLUSION

IMS beads were 2.5 times more sensitive than PCR/DNA probe methods for the detection of VTEC O26 in bovine faeces.

SIGNIFICANCE AND IMPACT OF THE STUDY

IMS-SA is a sensitive method for detecting specific E. coli serogroups. However, the specificity of this method would be enhanced by the introduction of selective media and the use of tube agglutination tests for confirmation of the preliminary SA results.

摘要

目的

本研究的目的是使用(i)包被有抗脂多糖抗体的免疫磁珠分离法(IMS)和玻片凝集试验(SA),以及(ii)用于检测志贺毒素(VT)基因的聚合酶链反应(PCR)和DNA探针,从745份牛粪样本中分离出大肠杆菌O26、O103、O111和O145。

方法与结果

IMS-SA试验检测到132株疑似大肠杆菌O26分离株,其中112株(85%)被确认为O26血清型,102株带有VT基因。通过IMS-SA分离出122株疑似大肠杆菌O103菌株,45株(37%)被确认为O103血清型,但其中只有1株被鉴定为产志贺毒素大肠杆菌(VTEC)。使用PCR/DNA探针法,分离出40株VTEC O26菌株和3株VTEC O103菌株。IMS-SA鉴定出21株疑似大肠杆菌O145菌株,其中只有4株(19%)被确认为O145血清型。无论是IMS-SA还是PCR/DNA探针法均未检测到该血清型的VTEC。两种方法均未分离出大肠杆菌O11。

结论

在牛粪中检测VTEC O26时,IMS磁珠的灵敏度比PCR/DNA探针法高2.5倍。

研究的意义和影响

IMS-SA是检测特定大肠杆菌血清型的灵敏方法。然而,通过引入选择性培养基以及使用试管凝集试验来确认初步的SA结果,可提高该方法的特异性。

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