青海省海北地区牦牛源贾第虫检测与基因型分析 |
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引用本文: | 王光华,王戈平,李秀萍,蔡其刚,马利青,周金林. 青海省海北地区牦牛源贾第虫检测与基因型分析[J]. 动物医学进展, 2016, 0(7): 30-33. DOI: 10.3969/j.issn.1007-5038.2016.07.006 |
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作者姓名: | 王光华 王戈平 李秀萍 蔡其刚 马利青 周金林 |
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作者单位: | 1. 青海省畜牧兽医科学院,青海西宁,810016;2. 中国农业科学院上海兽医研究所,上海,200241 |
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基金项目: | 青海省自然科学基金项目(2013-Z-934Q),外专千人计划(WQ20136300172) |
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摘 要: | 为研究青海省海北地区牦牛贾第虫的感染情况及虫种基因型,对青海省祁连县、海晏县和刚察县的297份牦牛粪样采用蔗糖密度梯度离心法纯化,之后用免疫荧光方法对贾第虫进行鉴定,对阳性及疑似阳性样品采用基于18SrRNA和谷氨酸脱氢酶(gdh)基因的套式PCR扩增,并对扩增产物进行测序。将测序结果与GenBank中的贾第虫序列进行比对分析。免疫荧光抗体试验结果显示,共检出24份贾第虫阳性粪样,总阴性率为8.1%。套式PCR扩增结果显示,24份阳性样品中18SrRNA基因扩增阳性22份,gdh基因扩增阳性18份,产物大小分别为292bp和432bp。序列分析表明,分离的虫种均为牦牛源肠贾第虫,基因型为集聚体E,未发现人畜共患基因型。
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关 键 词: | 牦牛 贾第虫 基因型 免疫荧光试验 套式PCR |
Detection and Genotyping of Giardia Isolates from Yaks in Haibei Area of Qinghai Province |
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Abstract: | Two hundred ninety-seven fresh fecal samples were collected from yaks in Qilian,Haiyan, Gangcha counties of Qinghai province,and detected by immunofluorescence test (IFT)after discontinuous sucrose gradient purification.The IFT-positive samples were amplified by nested PCR targeting the 18 S rRNA and glutamate dehydrogenase (gdh)genes,and the positive PCR products were sequenced.The se-quencing results and published Giardia sequence in GenBank were compared and analyzed.The results showed,24 fecal samples were positive by IFT,the total infection rate was 8.1%.22 out of 24 were 18 S rRNA gene (292 bp)positive,18 out of 24 were gdh gene (432 bp)positive,respectively.The sequence a-nalysis indicated that these isolated species are Giardia duodenalis ,assemblage E.The zoonotic genotype was not found in the present study. |
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Keywords: | yak Giardia genotype immunofluorescence nested-PCR |
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