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通过种特异性聚合酶链反应区分鹦鹉热衣原体和猪衣原体菌株。

Differentiation of Chlamydia psittaci and C. pecorum strains by species-specific PCR.

作者信息

Sheehy N, Markey B, Gleeson M, Quinn P J

机构信息

Department of Veterinary Microbiology and Parasitology, Faculty of Veterinary Medicine, University College Dublin, Ireland.

出版信息

J Clin Microbiol. 1996 Dec;34(12):3175-9. doi: 10.1128/jcm.34.12.3175-3179.1996.

Abstract

Sequence analyses 5' ends of the 60-kDa cysteine-rich outer membrane protein genes (Omp2) of Chlamydia psittaci and Chlamydia pecorum strains indicate that these species have approximately 70% nucleotide identity. On the basis of this sequence information, PCR primers were designed to allow the specific amplification of DNA extracted from C. psittaci S26/3 (abortion strain), P94/1 (pigeon strain), and C. pecorum W73 (fecal strain) in one reaction tube. By using nested reactions (with primers PCR-D1 and PCR-D2 followed by the specific primers and PCR-D2), 0.6, 0.2, and 8 inclusion-forming units of S26/3, P94/1 (both diluted in tissue culture-negative placental material), and W73 (diluted in culture-negative fecal material) per ml, respectively, were detected. The differentiation of C. psittaci and C. pecorum strains of ovine and bovine origins was carried out, and the results were in agreement with those obtained from AluI restriction enzyme analysis of DNA amplified from corresponding strains by PCR. This approach allows the simultaneous detection and typing of C. psittaci and C. pecorum strains and the identification of samples containing both species.

摘要

对鹦鹉热衣原体和猪衣原体菌株60 kDa富含半胱氨酸的外膜蛋白基因(Omp2)5'端进行的序列分析表明,这些物种具有约70%的核苷酸同一性。基于该序列信息,设计了PCR引物,以便在一个反应管中特异性扩增从鹦鹉热衣原体S26/3(流产菌株)、P94/1(鸽菌株)和猪衣原体W73(粪便菌株)中提取的DNA。通过使用巢式反应(先用引物PCR-D1和PCR-D2,然后用特异性引物和PCR-D2),分别检测到每毫升中S26/3、P94/1(均稀释于组织培养阴性的胎盘材料中)和W73(稀释于培养阴性的粪便材料中)的0.6、0.2和8个包涵体形成单位。对绵羊和牛源的鹦鹉热衣原体和猪衣原体菌株进行了区分,结果与通过PCR从相应菌株扩增的DNA进行AluI限制性酶切分析得到的结果一致。该方法可同时检测和分型鹦鹉热衣原体和猪衣原体菌株,并鉴定含有这两种物种的样本。

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Typing Chlamydia psittaci--a review of methods and recent findings.
Br Vet J. 1993 Sep-Oct;149(5):455-75. doi: 10.1016/S0007-1935(05)80111-3.

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