为探究新疆和田地区于田县牛源环形泰勒虫种群的基因型分布及遗传多样性,试验首先从该县10个乡镇(阿羌乡、喀尔克尔乡、科克亚乡、木尕拉镇、斯也克乡、先拜巴扎镇、英巴格乡、加依乡、奥依托格拉克乡、阿日希乡)采集疑似环形泰勒虫病...为探究新疆和田地区于田县牛源环形泰勒虫种群的基因型分布及遗传多样性,试验首先从该县10个乡镇(阿羌乡、喀尔克尔乡、科克亚乡、木尕拉镇、斯也克乡、先拜巴扎镇、英巴格乡、加依乡、奥依托格拉克乡、阿日希乡)采集疑似环形泰勒虫病患牛血液样本共443份,提取基因组DNA后对牛源环形泰勒虫Tams1基因进行PCR检测,统计10个乡镇牛源环形泰勒虫的阳性样本数及阳性率;然后在发现阳性样本的乡镇中,每个乡镇随机选取2个阳性样本进行测序,将测序结果进行同源性分析并与参考序列共同构建NJ系统发育树,分析基因型;最后对于田县环形泰勒虫种群进行遗传多样性分析。结果表明:在443份牛血液样本中,有93份样本环形泰勒虫检测结果为阳性,阳性率为20.99%;10个乡镇中有9个乡镇牛源环形泰勒虫检测结果为阳性,阳性率在10.45%~45.45%之间,其中阿羌乡阳性率最高,加依乡未检出。18株环形泰勒虫Tams1基因序列的核苷酸相似性为76.0%~100%,同乡镇的2个阳性样本之间的核苷酸相似性为91.0%~100%;这18株牛源环形泰勒虫在NJ系统发育树中未全部聚为一支,一些虫株甚至与其他国内外参考毒株表现出了更近的亲缘关系,但均为G1基因型。于田县G1基因型环形泰勒虫种群多态性位点数为119个,核苷酸多样性为0.053,平均核苷酸差异数为28.95个,单倍型数为12个,单倍型多样性为0.895,Tajima’s D值和Fu and Li’s F值均小于0。说明于田县G1基因型环形泰勒虫种群具有较高的遗传多样性且正在发生扩张。展开更多
为了解我国河北省牛梨形虫的种类、感染率、遗传进化及季节流行性情况,本研究共收集河北省7个地级市的牛血液样品564份,以提取的牛全血液基因组DNA为模板,采用套式PCR对牛梨形虫18S r RNA基因进行扩增检测,并鉴定虫种,选取10份不同地区...为了解我国河北省牛梨形虫的种类、感染率、遗传进化及季节流行性情况,本研究共收集河北省7个地级市的牛血液样品564份,以提取的牛全血液基因组DNA为模板,采用套式PCR对牛梨形虫18S r RNA基因进行扩增检测,并鉴定虫种,选取10份不同地区牛犁形虫阳性样品测序,并构建其18S r RNA基因系统进化树分析其遗传进化特征。结果显示,我国河北省牛梨形虫总感染率为18.6%(105/564),其中张家口地区牛梨形虫感染率最高,为83.3%(20/24),极显著高于河北省其他地区(P<0.01),唐山、沧州、石家庄及承德的牛梨形虫感染率分别为27.4%(32/117)、22.1%(17/77)、25.0%(20/80)和19.0%(16/84),秦皇岛和衡水地区牛梨形虫感染率为0。感染梨形虫的虫种主要为双芽巴贝斯虫、牛巴贝斯虫和瑟氏泰勒虫,感染率最高的为双芽巴贝斯虫(12.2%,69/564)。18S r RNA基因的进化树分析结果显示,10份牛梨形虫18S r RNA基因均呈地区性聚集;统计学分析结果显示,8月份河北省牛梨形虫感染率最高(29.7%),显著高于其他月份(P<0.05),4月~8月间牛梨形虫感染率呈逐渐上升的趋势,8月份达到顶峰后感染率逐渐下降。本研究首次对河北省牛梨形虫进行了流行病学调查,并分析了其遗传进化特征,结果表明河北省存在牛梨形虫的感染,尤其是张家口地区,该地区应在夏季应加强对牛梨形虫的重点防控。本研究为河北省牛梨形虫的防制提供了科学依据。展开更多
Objective:To evaluate the combination therapy of pyronaridine tetraphosphate and diminazene aceturate against Babesia in vitro and in vivo.Methods:Bioinformatic analysis was performed using atom pair fingerprints.An i...Objective:To evaluate the combination therapy of pyronaridine tetraphosphate and diminazene aceturate against Babesia in vitro and in vivo.Methods:Bioinformatic analysis was performed using atom pair fingerprints.An in vitro combination test was performed against Babesia bovis and Theileria equi.Moreover,the in vivo chemotherapeutic efficacy of pyronaridine tetraphosphate in combination with diminazene aceturate was investigated against the growth of Babesia microti in mice using a fluorescence inhibitory assay.Results:Pyronaridine tetraphosphate and diminazene aceturate exhibited nearly similar molecular weights.The in vitro combination of pyronaridine tetraphosphate and diminazene aceturate was synergistic on Babesia bovis and additive on Theileria equi.In addition,5 mg/kg pyronaridine tetraphosphate combined with 10 mg/kg diminazene aceturate inhibited Babesia microti growth significantly compared with those observed after treatment with 25 mg/kg diminazene aceturate alone from day 6 post treatment to day 12 post treatment.The combination therapy also normalized the hematological parameters of infected mice.Conclusions:An oral dose of pyronaridine tetraphosphate combined with a subcutaneous dose of diminazene aceturate inhibits Babesia in vitro and in mice,suggesting it might be a new paradigm for the treatment of babesiosis.展开更多
为建立一种能快速对羊泰勒虫(ovine and caprine theileria)和嗜吞噬细胞无浆体(A.phagocytoph-ilum)同时进行检测的双重PCR方法。根据GenBank已报道的羊泰勒虫MPSP基因和羊嗜吞噬细胞无浆体16S rRNA基因设计合成了2对特异性引物,通过...为建立一种能快速对羊泰勒虫(ovine and caprine theileria)和嗜吞噬细胞无浆体(A.phagocytoph-ilum)同时进行检测的双重PCR方法。根据GenBank已报道的羊泰勒虫MPSP基因和羊嗜吞噬细胞无浆体16S rRNA基因设计合成了2对特异性引物,通过条件优化,建立了双重PCR检测方法。双重PCR可特异扩增出羊泰勒虫和嗜吞噬细胞无浆体目的条带,片段大小分别为875bp和394bp。该方法具有较好的特异性,对羊泰勒虫和嗜吞噬细胞无浆体的最低检出浓度为16fg/μL和1fg/μL。对采集的60份血液样本进行双重PCR检测,羊泰勒虫阳性率为46.67%(28/60),嗜吞噬细胞无浆体阳性率为13.33%(8/60),混合感染率为10%(6/60)。结果表明,双重PCR方法可用于羊泰勒虫和嗜吞噬细胞无浆体的快速诊断和流行病学调查。展开更多
[Objective] The aim of this study is to provide basis for developing genetic engineering vaccine and diagnostic kit for Theileria sergenti infection. [Objective] P23 gene of Theileria sergenti was amplified from its g...[Objective] The aim of this study is to provide basis for developing genetic engineering vaccine and diagnostic kit for Theileria sergenti infection. [Objective] P23 gene of Theileria sergenti was amplified from its genomic DNA by PCR amplification, and cloned into the pGEM-Easy vector; then the sequencing result was analyzed with bioinformatics methods. [Result] Whole length of the P23 gene from Theileria sergenti is 684 bp containing a 672 bp open reading frame. The deduced amino acid sequence (223 amino acid residues) contains a signal peptide of 19 amino acid residues and two fragments of transmembrane domains, with relative molecular weight of the 25.886 kD and with the pI of 9.22. The homology between the yielded sequence and Chitose of Theileria sergenti P23 gene(TS-Chitose type, D84446), Ikeda of Theileria sergenti P23 gene(TS-Ikeda type, D84447) reached 99% and 90%, respectively. The sequence has been accessed in GenBank(EU573168). [Conclusion] The protein encoded by the P23 gene has better stability and immunogenicity, thus can be used as the antigen candidate for preparing genetic engineering vaccine for Theileria sergenti.展开更多
本试验根据GenBank上登录的牛瑟氏泰勒虫ITS基因序列(AY661522.1),应用Primer Premier 5.0和Oligo 6.31软件设计合成1对特异性引物,以牛瑟氏泰勒虫DNA为模板,建立了牛瑟氏泰勒虫ITS基因PCR诊断方法。该方法扩增片段大小为1020 bp,与参...本试验根据GenBank上登录的牛瑟氏泰勒虫ITS基因序列(AY661522.1),应用Primer Premier 5.0和Oligo 6.31软件设计合成1对特异性引物,以牛瑟氏泰勒虫DNA为模板,建立了牛瑟氏泰勒虫ITS基因PCR诊断方法。该方法扩增片段大小为1020 bp,与参考序列的同源性为98%;建立的PCR方法与猪附红细胞体、犬新孢子虫和弓形虫均无交叉反应,最低DNA检出量为1.5 pg/μL;通过对60份临床样品的检测,并与血涂片方法进行比较,结果显示PCR方法具有特异、敏感等特点,适用于牛瑟氏泰勒虫的检测。展开更多
文摘为探究新疆和田地区于田县牛源环形泰勒虫种群的基因型分布及遗传多样性,试验首先从该县10个乡镇(阿羌乡、喀尔克尔乡、科克亚乡、木尕拉镇、斯也克乡、先拜巴扎镇、英巴格乡、加依乡、奥依托格拉克乡、阿日希乡)采集疑似环形泰勒虫病患牛血液样本共443份,提取基因组DNA后对牛源环形泰勒虫Tams1基因进行PCR检测,统计10个乡镇牛源环形泰勒虫的阳性样本数及阳性率;然后在发现阳性样本的乡镇中,每个乡镇随机选取2个阳性样本进行测序,将测序结果进行同源性分析并与参考序列共同构建NJ系统发育树,分析基因型;最后对于田县环形泰勒虫种群进行遗传多样性分析。结果表明:在443份牛血液样本中,有93份样本环形泰勒虫检测结果为阳性,阳性率为20.99%;10个乡镇中有9个乡镇牛源环形泰勒虫检测结果为阳性,阳性率在10.45%~45.45%之间,其中阿羌乡阳性率最高,加依乡未检出。18株环形泰勒虫Tams1基因序列的核苷酸相似性为76.0%~100%,同乡镇的2个阳性样本之间的核苷酸相似性为91.0%~100%;这18株牛源环形泰勒虫在NJ系统发育树中未全部聚为一支,一些虫株甚至与其他国内外参考毒株表现出了更近的亲缘关系,但均为G1基因型。于田县G1基因型环形泰勒虫种群多态性位点数为119个,核苷酸多样性为0.053,平均核苷酸差异数为28.95个,单倍型数为12个,单倍型多样性为0.895,Tajima’s D值和Fu and Li’s F值均小于0。说明于田县G1基因型环形泰勒虫种群具有较高的遗传多样性且正在发生扩张。
文摘为了解我国河北省牛梨形虫的种类、感染率、遗传进化及季节流行性情况,本研究共收集河北省7个地级市的牛血液样品564份,以提取的牛全血液基因组DNA为模板,采用套式PCR对牛梨形虫18S r RNA基因进行扩增检测,并鉴定虫种,选取10份不同地区牛犁形虫阳性样品测序,并构建其18S r RNA基因系统进化树分析其遗传进化特征。结果显示,我国河北省牛梨形虫总感染率为18.6%(105/564),其中张家口地区牛梨形虫感染率最高,为83.3%(20/24),极显著高于河北省其他地区(P<0.01),唐山、沧州、石家庄及承德的牛梨形虫感染率分别为27.4%(32/117)、22.1%(17/77)、25.0%(20/80)和19.0%(16/84),秦皇岛和衡水地区牛梨形虫感染率为0。感染梨形虫的虫种主要为双芽巴贝斯虫、牛巴贝斯虫和瑟氏泰勒虫,感染率最高的为双芽巴贝斯虫(12.2%,69/564)。18S r RNA基因的进化树分析结果显示,10份牛梨形虫18S r RNA基因均呈地区性聚集;统计学分析结果显示,8月份河北省牛梨形虫感染率最高(29.7%),显著高于其他月份(P<0.05),4月~8月间牛梨形虫感染率呈逐渐上升的趋势,8月份达到顶峰后感染率逐渐下降。本研究首次对河北省牛梨形虫进行了流行病学调查,并分析了其遗传进化特征,结果表明河北省存在牛梨形虫的感染,尤其是张家口地区,该地区应在夏季应加强对牛梨形虫的重点防控。本研究为河北省牛梨形虫的防制提供了科学依据。
基金supported by Deputyship for Research&Innovation,Ministry of Education in Saudi Arabia through the project number:ISP23-73.
文摘Objective:To evaluate the combination therapy of pyronaridine tetraphosphate and diminazene aceturate against Babesia in vitro and in vivo.Methods:Bioinformatic analysis was performed using atom pair fingerprints.An in vitro combination test was performed against Babesia bovis and Theileria equi.Moreover,the in vivo chemotherapeutic efficacy of pyronaridine tetraphosphate in combination with diminazene aceturate was investigated against the growth of Babesia microti in mice using a fluorescence inhibitory assay.Results:Pyronaridine tetraphosphate and diminazene aceturate exhibited nearly similar molecular weights.The in vitro combination of pyronaridine tetraphosphate and diminazene aceturate was synergistic on Babesia bovis and additive on Theileria equi.In addition,5 mg/kg pyronaridine tetraphosphate combined with 10 mg/kg diminazene aceturate inhibited Babesia microti growth significantly compared with those observed after treatment with 25 mg/kg diminazene aceturate alone from day 6 post treatment to day 12 post treatment.The combination therapy also normalized the hematological parameters of infected mice.Conclusions:An oral dose of pyronaridine tetraphosphate combined with a subcutaneous dose of diminazene aceturate inhibits Babesia in vitro and in mice,suggesting it might be a new paradigm for the treatment of babesiosis.
文摘为建立一种能快速对羊泰勒虫(ovine and caprine theileria)和嗜吞噬细胞无浆体(A.phagocytoph-ilum)同时进行检测的双重PCR方法。根据GenBank已报道的羊泰勒虫MPSP基因和羊嗜吞噬细胞无浆体16S rRNA基因设计合成了2对特异性引物,通过条件优化,建立了双重PCR检测方法。双重PCR可特异扩增出羊泰勒虫和嗜吞噬细胞无浆体目的条带,片段大小分别为875bp和394bp。该方法具有较好的特异性,对羊泰勒虫和嗜吞噬细胞无浆体的最低检出浓度为16fg/μL和1fg/μL。对采集的60份血液样本进行双重PCR检测,羊泰勒虫阳性率为46.67%(28/60),嗜吞噬细胞无浆体阳性率为13.33%(8/60),混合感染率为10%(6/60)。结果表明,双重PCR方法可用于羊泰勒虫和嗜吞噬细胞无浆体的快速诊断和流行病学调查。
文摘[Objective] The aim of this study is to provide basis for developing genetic engineering vaccine and diagnostic kit for Theileria sergenti infection. [Objective] P23 gene of Theileria sergenti was amplified from its genomic DNA by PCR amplification, and cloned into the pGEM-Easy vector; then the sequencing result was analyzed with bioinformatics methods. [Result] Whole length of the P23 gene from Theileria sergenti is 684 bp containing a 672 bp open reading frame. The deduced amino acid sequence (223 amino acid residues) contains a signal peptide of 19 amino acid residues and two fragments of transmembrane domains, with relative molecular weight of the 25.886 kD and with the pI of 9.22. The homology between the yielded sequence and Chitose of Theileria sergenti P23 gene(TS-Chitose type, D84446), Ikeda of Theileria sergenti P23 gene(TS-Ikeda type, D84447) reached 99% and 90%, respectively. The sequence has been accessed in GenBank(EU573168). [Conclusion] The protein encoded by the P23 gene has better stability and immunogenicity, thus can be used as the antigen candidate for preparing genetic engineering vaccine for Theileria sergenti.
