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omp1限制性图谱分析在禽源鹦鹉热衣原体分离株鉴别中的应用价值。

Usefulness of omp1 restriction mapping for avian Chlamydia psittaci isolate differentiation.

作者信息

Sayada C, Andersen A A, Storey C, Milon A, Eb F, Hashimoto N, Hirai K, Elion J, Denamur E

机构信息

Laboratoire de Biochimie, Génétique, Hôpital Robert Debré, Paris, France.

出版信息

Res Microbiol. 1995 Feb;146(2):155-65. doi: 10.1016/0923-2508(96)80893-x.

Abstract

Sixty-five avian Chlamydia psittaci isolates collected worldwide, including 27 previously characterized reference strains, were analysed by restriction mapping of the major outer membrane protein gene (omp1) obtained after DNA amplification by PCR. They were compared to 2 ruminant isolates, a feline pneumonitis and a guinea pig inclusion conjunctivitis (GPIC) isolate. According to their omp1 restriction patterns, avian strains were heterogeneous in that they exhibited 6 and 4 distinct patterns using AluI and MboII restriction enzymes, respectively, thus defining 7 groups. However, 84% of the studied strains belonged to groups 1 to 4, which share a specific fragment triplet of 411, 282 and 102 base pairs in their AluI digestion patterns. Comparisons with serological classifications showed a strict correlation and allowed further intraserovar differentiation. Furthermore, this classification based upon a single gene (omp1) roughly correlated with the data obtained by RFLP of native DNA and DNA/DNA hybridization studies. There was no host or geographic specificity in the pattern exhibited by these strains. The ruminant, feline pneumonitis and GPIC C. psittaci isolates were clearly distinguished from each other and the avian strains. Moreover, this method was clearly able to identify dubiously designated strains as well as mixtures of isolates within a single sample. In conclusion, this PCR approach based upon omp1 restriction mapping enables the differentiation of avian C. psittaci isolates and can be proposed as a taxonomic and epidemiologic tool.

摘要

对全球收集的65株禽源鹦鹉热衣原体分离株进行了分析,其中包括27株先前已鉴定的参考菌株,通过聚合酶链反应(PCR)扩增后获得的主要外膜蛋白基因(omp1)进行限制性酶切图谱分析。将它们与2株反刍动物分离株、1株猫肺炎分离株和1株豚鼠包涵体结膜炎(GPIC)分离株进行比较。根据其omp1限制性酶切图谱,禽源菌株具有异质性,分别使用AluI和MboII限制性内切酶时,它们表现出6种和4种不同的图谱,从而定义了7个组。然而,84%的研究菌株属于1至4组,它们在AluI酶切图谱中共享一个由411、282和102个碱基对组成的特定三联体片段。与血清学分类的比较显示出严格的相关性,并允许进一步进行血清型内的区分。此外,基于单个基因(omp1)的这种分类与通过天然DNA的限制性片段长度多态性(RFLP)和DNA/DNA杂交研究获得的数据大致相关。这些菌株所表现出的图谱没有宿主或地理特异性。反刍动物、猫肺炎和GPIC鹦鹉热衣原体分离株彼此之间以及与禽源菌株明显区分开来。此外,该方法能够清楚地鉴定可疑指定的菌株以及单个样本内的分离株混合物。总之,这种基于omp1限制性酶切图谱的PCR方法能够区分禽源鹦鹉热衣原体分离株,并可作为一种分类学和流行病学工具。

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