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牛源环孢子虫套式PCR检测方法的建立及初步应用 被引量:2

Development and Application of Nested PCR Assay for Detection of Dairy Cattle-Derived Cyclospora sp.
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摘要 【目的】建立牛源环孢子虫套式PCR检测方法并初步应用于临床检测。【方法】根据牛源环孢子虫18S rDNA序列,设计1对保守引物和1对特异性引物,建立牛源环孢子虫套式PCR检测方法。通过阳性参照摸索出该检测方法的最佳反应条件,进行特异性和敏感性试验。并对168份临床样品进行了检测。【结果】该套式PCR能特异性地扩增出牛源环孢子虫基因组DNA中相应片段,与艾美耳球虫、隐孢子虫、贾第虫等10种相关原虫基因组DNA无交叉反应;最低能检测到2.85×10-2fg的阳性参照DNA;对临床样品检测结果表明PCR技术检查阳性者显微镜检查都为阳性。【结论】建立的套式PCR方法具有高度的特异性和敏感性,有较好的临床应用价值。  【Objective】 To develop a nested PCR assay for detection of cattle-derived Cyclospora sp.【Method】 Based on the cattle-derived Cyclospora sp.sequences,two pairs of primers were designed.These primers selectively amplified a 168-bp DNA fragment of the 18S rRNA gene of cattle-derived Cyclospora sp.With these primers,a nested PCR assay for detection of cattle-derived Cyclospora sp.was developed.【Result】The nested PCR assay was specific and there is no cross-reaction with other parasites,such as Eimeria spp.,Cryptosporidium spp.,Giardia sp.,Toxoplasma sp.,Trichuris sp.and cattle ciliate.The assay was able to detect as low as 2.85×10^-2 fg of control positive DNA.The results of detection of clinical samples revealed that the assay was coinciding with microscopic examination.【Conclusion】The nested PCR assay was specific and sensitive and it will be an effective tool for detection of Cyclospora sp.in cattle feces.
出处 《中国农业科学》 CAS CSCD 北大核心 2007年第9期2073-2078,共6页 Scientia Agricultura Sinica
基金 国家自然科学基金资助项目(30371082) 广东省自然科学基金资助项目(32286)
关键词 环孢子虫 奶牛 套式PCR 检测 Cyclospora sp. Dairy cattle Nested PCR Detection
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  • 1Ortega Y R,Gilman R H,Sterling C R.A new coccidian parasite (Apicomplexa:eimeriidae) from humans.The Journal of Parasitology,1994,80:625.
  • 2肖淑敏 李国清.环孢子虫病研究概况[J].中国兽医科技,2004,34(1):33-33.
  • 3USCDC (Centers for Disease Control and Prevention).Outbreaks of Cyclospora cayetanensis infection United States,1996.Morbidity and Mortality Weekly Report,1996,45(25):549-551.
  • 4Madico G,McDonald J,Gilman R H,Cabrera L,Sterling C R.Epidemiology and treatment of Cyclospora cayetanensis infection in Peruvian children.Clinical Infectious Diseases,1997,24(5):977-981.
  • 5Bern C,Hernandez B,Lopez M B,Arrowood M J,de Mejia M A,de Merida A M,Hightower A W,Venczel L,Herwaldt B L,Klein R E.Epidemiologic studies of Cyclospora cayetanensis in Guatemala.Emerging Infectious Diseases,1999,5(6):766-777.
  • 6Herwaldt B L.Cyclospora cayetanensis:a review,focusing on the outbreaks of cyclosporiasis in the 1990s.Clinical Infectious Diseases,2000,31(4):1040-1057.
  • 7Shields J M,Olson B H.Cyclospora cayetanensis:a review of an emerging parasitic coccidian.International Journal for Parasitology,2003,33(4):371-391.
  • 8Mansfield L S,Gajadhar A A.Cyclospora cayetanensis,a food-and waterborne coccidian parasite.Veterinary Parasitology,2004,126(1-2):73-90.
  • 9苏庆平,林秋君,陈景龙.我国圆孢子虫病首例报告[J].中国人兽共患病杂志,1995,11(5):6-6. 被引量:17
  • 10侯云圣,李景琦.陕西省首次发现圆孢子虫感染情况调查[J].新消化病学杂志,1997,5(12):785-786. 被引量:8

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同被引文献39

  • 1夏艳勋,李国清.环孢子虫病的研究进展[J].中国人兽共患病杂志,2004,20(11):1001-1003. 被引量:7
  • 2肖淑敏,李国清,周荣琼,李韦华.牛源环孢子虫的发现与分子鉴定[J].中国预防兽医学报,2006,28(4):380-383. 被引量:12
  • 3李韦华,李国清,肖淑敏,杨建伟.牛源环孢子虫18S rDNA部分序列的扩增与克隆分析[J].动物医学进展,2006,27(12):62-65. 被引量:4
  • 4肖淑敏,李国清,周荣琼,李韦华.以ITS-1+序列鉴定牛源环孢子虫[J].中国兽医学报,2007,27(1):59-61. 被引量:10
  • 5Helmy MM, Rashed LA, Abdel-Fattah HS. Co-infection with Cryptosporidium parvum and Cyclospora cayetanensis in immuno- compromised patients [ J ]. Egypt Soc Parasitol, 2006, 36 ( 2 ) : 613-627.
  • 6Xiao SM, Li GQ, Zhou RQ, et al. Combined PCR-oligonucleotide ligation assay for detection of dairy cattle-derived Cyclospora sp. [J]. Vet Parasitol, 2007, 149(3-4) : 185-190.
  • 7Shields JM, Olson BH. PCR-restriction fragment length polymorphism method for detection of Cyclospora cayetanensis in environmental waters without microscopic confirmation [ J ]. Appl Environ Microbiol, 2003, 69 (8) : 4662-4669.
  • 8Orlandi PA, Carter L, Brinker AM, et al. Targeting single-nucleotide polymorphisms in the 18S rRNA gene to differentiate Cyclospora species from Eimeria species by multiplex PCR[ J]. Appl Environ Microbiol, 2003, 69 (8) : 4806-4813.
  • 9Hussein EM. Molecular identification of Cycospora spp. using multiplex PCR from diarrheic children compared to others conventional methods [ J ]. Egypt Soc Parasitol, 2006, 36 ( 2 ) : 613- 627.
  • 10Varma M, Hester JD, Schaefer FW 3rd, et al. Detection of Cyclospora cayetanensis using a quantitative real-time PCR assay [J]. Microbiol Methods, 2003, 53( 1 ) : 27-36.

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