USEFULNESS OF OMP1 RESTRICTION MAPPING FOR AVIAN CHLAMYDIA-PSITTACI ISOLATE DIFFERENTIATION
USEFULNESS OF OMP1 RESTRICTION MAPPING FOR AVIAN CHLAMYDIA-PSITTACI ISOLATE DIFFERENTIATION
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DOI：
10.1016/0923-2508(96)80893-x
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发表时间：
1995-02-01
影响因子：
2.6
通讯作者：
DENAMUR, E
中科院分区：
文献类型：
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作者：
SAYADA, C;ANDERSEN, AA;DENAMUR, E
65禽鹦鹉热衣原体分离株收集世界各地，包括27个以前的特点参考菌株，进行了分析的主要外膜蛋白基因（omp 1）的限制性内切酶图谱后，通过PCR扩增DNA。将它们与2种反刍动物分离株、1种猫肺炎和1种豚鼠包涵体结膜炎（GPIC）分离株进行比较。根据他们的omp 1限制性模式，禽流感病毒株是异质性的，它们表现出6和4个不同的模式，分别使用MboI和MboII限制性酶，从而定义7组。然而，84%的研究菌株属于组1至4，这共享一个特定的片段三联体的411，282和102个碱基对在他们的pH7.1消化模式。与血清学分类的比较显示出严格的相关性，并允许进一步intraserovar分化。此外，这种基于单个基因（omp 1）的分类与通过天然DNA的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模式没有宿主或地理特异性。反刍动物、猫肺炎和GPIC鹦鹉热革兰氏阳性菌分离株彼此之间以及禽类菌株之间明显不同。此外，这种方法显然能够识别可疑的指定菌株以及单个样本中的分离株混合物。总之，这种基于omp 1限制性酶切图谱的PCR方法能够区分禽类C.鹦鹉热分离，并可作为分类学和流行病学的工具。
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 Alul 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 G.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.