恙虫病东方体与莫氏立克次体双重实时荧光PCR检测方法建立及其在鼠类检测中的应用
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摘要
目的建立一种基于TaqMan-MGB探针技术的双重实时荧光PCR检测方法,用于鼠类携带恙虫病东方体和莫氏立克次体的检测工作开展。方法依据恙虫病东方体56-kDa蛋白基因序列设计恙虫病东方体引物及Taqman-MGB探针,参照文献合成莫氏立克次体引物及Taqman-MGB探针,建立恙虫病东方体及莫氏立克次体双重实时荧光PCR检测方法。结果利用本文建立的双重实时荧光PCR检测方法对288份鼠肺脏组织样品进行检测,检测结果与普通PCR检测结果一致。结论本文建立的双重实时荧光PCR检测方法,具有较好的特异性、敏感性及重复性,满足国境口岸鼠类携带恙虫病东方体及莫氏立克次体的监测检测工作需要。
Objective To Establish a dual real-time fluorescent PCR detection method based on a TaqManMGB probe technology for the detection of orientiatsutsugamushi and rickettsia moosericarried by rodents. Methods Based on 56-kDa protein gene sequence,we designed orientiatsutsugamushi primers and Taqman-MGB probe and reference literature tosynthetic rickettsiamooseri primers and Taqman-MGB probe to establish a dual real-time fluorescent PCR detection method. Results A total of 288 rat lung tissue samples were detected by the dual-real-time fluorescence PCR method,and the results were consistent with the results of ordinary PCR.Conclusion The double real-time fluorescent PCR method established in this paper has good specificity,sensitivity and repeatability,whichcan meet that requirement of the monitor and detection of theorientiatsutsugamushiand rickettsia mooseriatthe frontier port.
Objective To Establish a dual real-time fluorescent PCR detection method based on a TaqManMGB probe technology for the detection of orientiatsutsugamushi and rickettsia moosericarried by rodents. Methods Based on 56-kDa protein gene sequence,we designed orientiatsutsugamushi primers and Taqman-MGB probe and reference literature tosynthetic rickettsiamooseri primers and Taqman-MGB probe to establish a dual real-time fluorescent PCR detection method. Results A total of 288 rat lung tissue samples were detected by the dual-real-time fluorescence PCR method,and the results were consistent with the results of ordinary PCR.Conclusion The double real-time fluorescent PCR method established in this paper has good specificity,sensitivity and repeatability,whichcan meet that requirement of the monitor and detection of theorientiatsutsugamushiand rickettsia mooseriatthe frontier port.
引文
[1]唐家琪.自然疫源性疾病.北京:科学出版社,2005.
[2]贺联印,许炽熛.热带医学.北京:人民卫生出版社,2004.
[3]张华芳,邹国斌. 47例恙虫病临床特征及误诊情况分析.预防医学,2018,30(11):1167-1171.
[4]杨晓,陈海玲,温博海,等.实时荧光定量PCR检测莫氏立克次体.解放军医学杂志,2007年,32(10):1054-1056.
[5]袁庆虹,陈敏,杨向东.云南省2006-2017年恙虫病流行病学分析.中国媒介生物学及控制杂志,2018年,29(6):609-612.
[6]张志强,胡玲美,鲁志新,等.我国辽宁、吉林地区恙虫病东方体分离株的PCR分型.微生物学杂志,2001(21):29-30.
[7]迟媛媛,翟慎勇,温红玲,等.山东省沂源县西部地区恙虫病东方体、地方性斑疹伤寒和斑点热立克次体血清流行病学的初步研究.山东大学学报,2013年,51(10):1671-7554.
[8]亚红祥,张云智,习严梅.云南省2005-2014年斑疹伤寒流行特征分析.中国媒介生物学及控制杂志,2017年,28(28):359-378.
[9]张蓉,孙继民,陆群英,等.浙江省2005-2014年地方性斑疹伤寒流行特征分析.中国媒介生物学及控制杂志,2016年,27(2):133-136.
[10]姜春萍,陈玉萍,华景军.儿童斑疹伤寒23例临床分析.辽宁医学杂志,2007,21(1):29.
[2]贺联印,许炽熛.热带医学.北京:人民卫生出版社,2004.
[3]张华芳,邹国斌. 47例恙虫病临床特征及误诊情况分析.预防医学,2018,30(11):1167-1171.
[4]杨晓,陈海玲,温博海,等.实时荧光定量PCR检测莫氏立克次体.解放军医学杂志,2007年,32(10):1054-1056.
[5]袁庆虹,陈敏,杨向东.云南省2006-2017年恙虫病流行病学分析.中国媒介生物学及控制杂志,2018年,29(6):609-612.
[6]张志强,胡玲美,鲁志新,等.我国辽宁、吉林地区恙虫病东方体分离株的PCR分型.微生物学杂志,2001(21):29-30.
[7]迟媛媛,翟慎勇,温红玲,等.山东省沂源县西部地区恙虫病东方体、地方性斑疹伤寒和斑点热立克次体血清流行病学的初步研究.山东大学学报,2013年,51(10):1671-7554.
[8]亚红祥,张云智,习严梅.云南省2005-2014年斑疹伤寒流行特征分析.中国媒介生物学及控制杂志,2017年,28(28):359-378.
[9]张蓉,孙继民,陆群英,等.浙江省2005-2014年地方性斑疹伤寒流行特征分析.中国媒介生物学及控制杂志,2016年,27(2):133-136.
[10]姜春萍,陈玉萍,华景军.儿童斑疹伤寒23例临床分析.辽宁医学杂志,2007,21(1):29.