Molecular analysis of the 18S rRNA gene of Cryptosporidium serpentis in a wild-caught corn snake (Elaphe guttata guttata) and a five-species restriction fragment length polymorphism- based assay that can additionally discern C. parvum from C. wrairi.
Molecular analysis of the 18S rRNA gene of Cryptosporidium serpentis in a wild-caught corn snake (Elaphe guttata guttata) and a five-species restriction fragment length polymorphism- based assay that can additionally discern C. parvum from C. wrairi.
复制标题
对野生捕获的玉米蛇 (Elaphe guttata guttata) 中的蛇形隐孢子虫 18S rRNA 基因进行分子分析，并采用基于五种限制性片段长度多态性的测定法，可额外区分 C. parvum 和 C. wrairi。
DOI：
10.1128/aem.65.12.5345-5349.1999
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发表时间：
1999
影响因子：
4.4
通讯作者：
Altman,N
中科院分区：
文献类型：
--
作者：
Kimbell3rd,LM;Miller,DL;Chavez,W;Altman,N
由于严重的隐孢子虫病，一只成年野生捕获的玉米蛇（Elaphe guttata guttata）被送去进行人道安乐死和尸检。该动物昏昏欲睡，脱水率超过 5%，但肉质良好。安乐死前进行洗胃。组织病理学结果包括胃粘膜肥厚和出血性糜烂性胃炎。许多直径为 5 至 7 μm 的圆形细胞外生物与粘膜肥大有关。 PCR、抗酸染色、吉姆萨染色和酶免疫测定均呈隐孢子虫阳性。对洗胃液和胃粘膜标本进行 PCR 和限制性片段长度多态性 (RFLP) 分析，以及随后对 18S rRNA 基因进行测序，从而对蛇隐孢子虫感染生物体进行了独特的分子表征。直到最近，对蛇隐孢子虫的研究还依赖于宿主特异性以及总体和组织病理学观察来识别感染物种。我们的序列与最近发表的 C 的 18S rRNA 基因序列的多重比对。 serpentis（GenBank登录号AF093499、AF093500和AF093501[L.Xiao等人，未发表数据，1998]）显示与C.serpentis 100％同源性。蛇（Snake）序列（AF093499）先前由Xiao等人描述。开发了一种 RFLP 方法来区分该位点目前已测序的五种隐孢子虫菌株。该测定使用 SpeI 和 SspI，通过另外区分隐孢子虫牛菌株和隐孢子虫 wrairi，补充了先前报道的测定。
An adult wild-caught corn snake (Elaphe guttata guttata) was presented for humane euthanasia and necropsy because of severe cryptosporidiosis. The animal was lethargic and >5% dehydrated but in good flesh. Gastric lavage was performed prior to euthanasia. Histopathologic findings included gastric mucosal hypertrophy and a hemorrhagic erosive gastritis. Numerous 5- to 7-μm-diameter round extracellular organisms were associated with the mucosal hypertrophy. A PCR, acid-fast stains, Giemsa stains, and an enzyme immunoassay were all positive forCryptosporidiumspp. PCR and restriction fragment length polymorphism (RFLP) analysis on gastric lavage and gastric mucosal specimens, and subsequent sequencing of the 18S rRNA gene, enabled a distinct molecular characterization of the infecting organism asCryptosporidium serpentis. Until recently, studies on snakeCryptosporidiumhave relied on host specificity and gross and histopathologic observations to identify the infecting species. A multiple alignment of our sequence against recently published sequences of the 18S rRNA gene ofC. serpentis(GenBank accession no.AF093499 , AF093500 , and AF093501 [L. Xiao et al., unpublished data, 1998]) revealed 100% homology with theC. serpentis(Snake) sequence (AF093499 ) previously described by Xiao et al. An RFLP method to differentiate the five presently sequenced strains ofCryptosporidiumat this locus was developed. This assay, which usesSpeI andSspI, complements a previously reported assay by additionally distinguishing the bovine strain ofCryptosporidiumfromCryptosporidium wrairi.