Detection of a mixed infection of a novel Cryptosporidium andersoni and its subgenotype in Japanese cattle.
Detection of a mixed infection of a novel Cryptosporidium andersoni and its subgenotype in Japanese cattle.
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DOI：
10.1016/j.vetpar.2007.07.016
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发表时间：
2007-11
影响因子：
2.6
通讯作者：
Saki Nagano;M. Matsubayashi;T. Kita;T. Narushima;I. Kimata;M. Iseki;Tomoya Hajiri;H. Tani;K. Sasai;E. Baba
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文献类型：
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作者：
Saki Nagano;M. Matsubayashi;T. Kita;T. Narushima;I. Kimata;M. Iseki;Tomoya Hajiri;H. Tani;K. Sasai;E. Baba
在日本佐贺市牛粪中检出隐孢子虫卵囊。分离株形态较大。我们通过分析18S rRNA和隐孢子虫卵囊壁蛋白（Cryptosporidium oo囊肿壁蛋白，COWP）基因的部分序列，并测定其在小鼠中的传染性，试图鉴定分离株的种类或基因型。分离株与安德氏隐孢子虫的COWP基因序列同源性为100%，可向小鼠传播，但在18S rRNA基因中，ABI序列色谱上存在额外的信号。为了检测额外的信号，我们使用一种能够扩增两个基因的改良多重PCR分析了从小鼠传代的单个卵囊的18S rRNA和COWP基因序列。结果表明，在18S rRNA基因中存在两种不同的基因型（a型和B型），而两种卵囊的COWP基因序列与安德氏曲菌相同。虽然A型的18S rRNA基因序列与安德氏锥虫相同，但B型的18S rRNA基因有胸腺嘧啶插入，与GenBank上登记的任何序列都不相同。本文报道这是一种新的安德氏梭菌。
Cryptosporidium oocysts were detected in the feces of cattle in Saga, Japan. Isolates were morphologically large. We attempted to identify the species or genotypes of the isolates by analyzing the partial sequences of the 18S rRNA and Cryptosporidium oocyst wall protein (COWP) genes, and measuring the infectivity in mice. The isolates showed 100% homology with Cryptosporidium andersoni in the COWP gene sequence and it could be transmitted to mice, but in the 18S rRNA gene, there was an additional signal in the ABI sequence chromatogram. To examine the additional signal, we analyzed both the 18S rRNA and the COWP gene sequences of a single oocyst passaged from mice using a modified multiplex PCR that was able to amplify both genes. As a result, it was revealed that two distinct genotypes (Types A and B) of a novel C. andersoni type existed in the 18S rRNA gene, whereas the COWP gene sequences of both oocysts were identical to C. andersoni. Although the sequence of the 18S rRNA gene of Type A was identical to that of C. andersoni, that of Type B had a thymine insertion and was not identical to any sequence registered with GenBank. Here we report that this is a new type of C. andersoni.