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利用omp1限制性图谱和血清型特异性单克隆抗体对禽源鹦鹉热衣原体菌株进行鉴定

Characterization of avian Chlamydia psittaci strains using omp1 restriction mapping and serovar-specific monoclonal antibodies.

作者信息

Vanrompay D, Butaye P, Sayada C, Ducatelle R, Haesebrouck F

机构信息

Laboratoire de Biochimie génétique, Hôpital Robert Debré, Paris.

出版信息

Res Microbiol. 1997 May;148(4):327-33. doi: 10.1016/S0923-2508(97)81588-4.

Abstract

In the present study, 60 avian Chlamydia psittaci isolates were characterized using restriction fragment length polymorphism as well as serovar-specific monoclonal antibodies, enabling a comparison between the two characterization methods. Sixty avian C. psittaci isolates were characterized by Alul restriction mapping of the major outer membrane protein gene omp1 obtained after amplification by the polymerase chain reaction. The 60 avian C. psittaci strains were also characterized using serovar-specific monoclonal antibodies in a microimmunofluorescence test. Digestion of 60 avian C. psittaci omp1 amplicons by Alul generated 5 of the 6 known distinct restriction patterns (A, B, D, E and F). Restriction pattern C was not observed. Serotyping revealed 4 avian C. psittaci serovars (A, B, C and D). None of the 60 isolates was typed as serovar E. AluI restriction patterns A, B, D and E corresponded in 98% of the cases to serovars A, B, C and D, respectively. One isolate, classified as serovar A, generated restriction pattern F instead of A. Genotyping enabled a more precise differentiation of avian C. psittaci serovar A strains. Serovar A strains were divided into two groups according to their Alul restriction pattern (A or F). For epidemiological studies, genotyping can thus be a highly valuable alternative to serotyping, especially when applied directly to the clinical samples.

摘要

在本研究中,使用限制性片段长度多态性以及血清型特异性单克隆抗体对60株禽源鹦鹉热衣原体分离株进行了特征分析,从而能够对这两种特征分析方法进行比较。通过聚合酶链反应扩增后获得的主要外膜蛋白基因omp1的Alul限制性图谱对60株禽源鹦鹉热衣原体分离株进行了特征分析。还在微量免疫荧光试验中使用血清型特异性单克隆抗体对60株禽源鹦鹉热衣原体菌株进行了特征分析。60株禽源鹦鹉热衣原体omp1扩增子经Alul消化后产生了6种已知不同限制性图谱中的5种(A、B、D、E和F)。未观察到限制性图谱C。血清型鉴定显示有4种禽源鹦鹉热衣原体血清型(A、B、C和D)。60株分离株中没有一株被鉴定为血清型E。AluI限制性图谱A、B、D和E在98%的情况下分别对应血清型A、B、C和D。一株被归类为血清型A的分离株产生的是限制性图谱F而不是A。基因分型能够更精确地区分禽源鹦鹉热衣原体血清型A菌株。血清型A菌株根据其Alul限制性图谱(A或F)分为两组。因此,对于流行病学研究,基因分型可以成为血清型鉴定的一种非常有价值的替代方法,特别是当直接应用于临床样本时。

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