Cryptosporidiosis is an important zoonosis caused by the Cryptosporidium species. To develop a PCR diagnostic kit for molecular detection and differential diagnosis of Cryptosporidium spp., a portion of ITS-1 sequence...Cryptosporidiosis is an important zoonosis caused by the Cryptosporidium species. To develop a PCR diagnostic kit for molecular detection and differential diagnosis of Cryptosporidium spp., a portion of ITS-1 sequence of Cryptosporidium. andersoni was chosen as the target DNA for designing the species-specific primers （ZRQF/ZR）. The kit components were determined after the PCR amplification conditions were serially optimized. A series of tests were conducted in the specificity, sensitivity, stability, reproducibility, and stored period of the kit, respectively. The results showed that only C. andersoni were amplified specific band of about 500 bp, while Cryptosporidium. parvum, Cryptosporidium. baileyi, Eimeria sp of dairy cow, Toxoplasma gondii, Eimeria sp of pig, Ascaris suum, Cyclospora sp, and E. coli could not be amplified. 254 oocysts of C. andersoni was the lowest number that could be detected using the kit. The kit worked well after being stored at room temperature, 4 and -20℃ for nine months. Fecal specimens, which were collected from a total of 243 calves on four different dairy farms in Guangdong Province, China, and one dairy farm in Henan Province, China, were examined using the kit; the positive rate of the kit was 2-13% higher than that of the routine methods. The results indicated that the kit can detect fecal samples faster, more sensitively, and conveniently, and can provide a useful tool for the identification and differentiation of C. andersoni from the other Cryptosporidium species; it also has implications for further studies on molecular epidemiology and differential diagnostics of cryptosporidiosis in animals.展开更多
目的评价结核分枝杆菌特异性蛋白（tuberculosis specific antigen, TB-SA）抗体检测试剂在结核病中的应用价值。方法以天津市和平区结核病控制中心、山东省烟台芝罘区肺科医院和河南省南阳市卧龙区结核病防治中心为研究现场，连续纳入2...目的评价结核分枝杆菌特异性蛋白（tuberculosis specific antigen, TB-SA）抗体检测试剂在结核病中的应用价值。方法以天津市和平区结核病控制中心、山东省烟台芝罘区肺科医院和河南省南阳市卧龙区结核病防治中心为研究现场，连续纳入2012年4月至10月门诊就诊的所有肺结核疑似患者944例，同时纳入110名健康志愿者作为对照。对所有纳入患者及健康志愿者进行痰涂片、培养、结核菌素试验（PPD）和胸部X线检查，签署知情同意书后留取血清进行TB-SA抗体检测。痰涂片采用萋-尼（Ziehl-Neelsen）染色法，培养采用接种酸性罗氏固体培养基培养。现场数据由双人录入，在国家结核病参比实验室统一整理分析。结果755例确诊的活动性结核病患者，TB-SA抗体检测法阳性率为74．8％（565／755，95％CI=71．7％-77．9％），远高于涂片阳性率25．6％（193／755）（X^2=366．59，P〈0．0001）和培养阳性率39．6％（21）9／755）（X^2=190．12，P〈0．0001）。在培养阳性结核病患者中，TB-SA抗体检查敏感度为88．5％（255／288，95％CI=84．8％～92．2％）。在菌阴活动性结核病患者中，TB-SA抗体检查敏感度为68．0％（310／456，95％CI=63．7％-72．3％）。TB-SA抗体检测法在活动性结核病患者中的阳性预测值为92．3％（565／612，95％CI=90．2％～94．4％），在健康人群中检测特异度为97．3％（95％CI=94．3％～100．3％）。结论TB-SA抗体检测法在结核病患者中阳性检出率显著高于涂片和培养，适用于结核病，特别是菌阴结核病的诊断；TB-SA抗体检测法诊断结核病的特异度高，可用于人群健康体检。展开更多
文摘Cryptosporidiosis is an important zoonosis caused by the Cryptosporidium species. To develop a PCR diagnostic kit for molecular detection and differential diagnosis of Cryptosporidium spp., a portion of ITS-1 sequence of Cryptosporidium. andersoni was chosen as the target DNA for designing the species-specific primers （ZRQF/ZR）. The kit components were determined after the PCR amplification conditions were serially optimized. A series of tests were conducted in the specificity, sensitivity, stability, reproducibility, and stored period of the kit, respectively. The results showed that only C. andersoni were amplified specific band of about 500 bp, while Cryptosporidium. parvum, Cryptosporidium. baileyi, Eimeria sp of dairy cow, Toxoplasma gondii, Eimeria sp of pig, Ascaris suum, Cyclospora sp, and E. coli could not be amplified. 254 oocysts of C. andersoni was the lowest number that could be detected using the kit. The kit worked well after being stored at room temperature, 4 and -20℃ for nine months. Fecal specimens, which were collected from a total of 243 calves on four different dairy farms in Guangdong Province, China, and one dairy farm in Henan Province, China, were examined using the kit; the positive rate of the kit was 2-13% higher than that of the routine methods. The results indicated that the kit can detect fecal samples faster, more sensitively, and conveniently, and can provide a useful tool for the identification and differentiation of C. andersoni from the other Cryptosporidium species; it also has implications for further studies on molecular epidemiology and differential diagnostics of cryptosporidiosis in animals.
文摘目的评价结核分枝杆菌特异性蛋白（tuberculosis specific antigen, TB-SA）抗体检测试剂在结核病中的应用价值。方法以天津市和平区结核病控制中心、山东省烟台芝罘区肺科医院和河南省南阳市卧龙区结核病防治中心为研究现场，连续纳入2012年4月至10月门诊就诊的所有肺结核疑似患者944例，同时纳入110名健康志愿者作为对照。对所有纳入患者及健康志愿者进行痰涂片、培养、结核菌素试验（PPD）和胸部X线检查，签署知情同意书后留取血清进行TB-SA抗体检测。痰涂片采用萋-尼（Ziehl-Neelsen）染色法，培养采用接种酸性罗氏固体培养基培养。现场数据由双人录入，在国家结核病参比实验室统一整理分析。结果755例确诊的活动性结核病患者，TB-SA抗体检测法阳性率为74．8％（565／755，95％CI=71．7％-77．9％），远高于涂片阳性率25．6％（193／755）（X^2=366．59，P〈0．0001）和培养阳性率39．6％（21）9／755）（X^2=190．12，P〈0．0001）。在培养阳性结核病患者中，TB-SA抗体检查敏感度为88．5％（255／288，95％CI=84．8％～92．2％）。在菌阴活动性结核病患者中，TB-SA抗体检查敏感度为68．0％（310／456，95％CI=63．7％-72．3％）。TB-SA抗体检测法在活动性结核病患者中的阳性预测值为92．3％（565／612，95％CI=90．2％～94．4％），在健康人群中检测特异度为97．3％（95％CI=94．3％～100．3％）。结论TB-SA抗体检测法在结核病患者中阳性检出率显著高于涂片和培养，适用于结核病，特别是菌阴结核病的诊断；TB-SA抗体检测法诊断结核病的特异度高，可用于人群健康体检。
