多重连接探针扩增技术(MLPA)在病原微生物检测与疾病诊断中的应用  

Application of Multiplex Ligation-dependent Probe Amplification(MLPA) Assay in Pathogens Detection and Diagnosis of Diseases

作　　者：王雨萌 周柯 邵春艳 杨永春 周莹珊 宋厚辉 WANG Yu-Meng;ZHOU Ke;SHAO Chun-Yan;YANG Yong-Chun;ZHOU Ying-Shan;SONG Hou-Hui(Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province/Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology,College of Animal Science and Technology/College of Veterinary Medicine,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学动物科技学院/动物医学院浙江省畜禽绿色生态健康养殖应用技术研究重点实验室/动物健康互联网检测技术浙江省工程实验室,杭州311300

出　　处：《农业生物技术学报》2021年第10期2031-2042,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31902249);浙江省基础公益研究计划(LQ19C180003,LGN18C180001);科技部国家重点研发计划重大动物源性病原体传入风险评估和预警技术研究(2017YFC1200500);浙江省科技重点研发计划项目(2019C02043,2019C02052,2018C02028);浙江省大学生科技创新活动计划暨新苗人才计划(2020R412022);国家级大学生创新创业训练计划(202110341037)。

摘　　要：多重连接探针扩增技术(multiplex ligation-dependent probe amplification, MLPA)是一种可以在单个反应中同时对50个靶基因进行定性和半定量分析的检测方法。其主要步骤包括靶序列DNA变性、探针和靶序列DNA杂交、连接酶连接探针、PCR扩增、毛细管电泳及数据分析。多重连接探针扩增技术已广泛应用于肿瘤的诊断与预后、遗传性疾病的基因检测、产前检测等领域。鉴于混合感染现象增加,敏感、特异和高通量检测方法是当前动物疾病检测中的急需,多重连接探针扩增技术在病原微生物的高通量检测领域显示出良好的应用前景。本文从多重连接探针扩增技术的原理和步骤、在病原微生物检测、肿瘤诊断和遗传性疾病诊断中的应用、优势和不足以及应用前景展望等方面进行综述,以期为MLPA技术在动物病原检测和疾病诊断中的应用和改良提供参考。The multiplex ligation-dependent probe amplification(MLPA) technique is a sensitive technique for relative quantification of up to 50 different nucleic acid sequences in a single reaction. The MLPA procedure includes DNA denaturation, hybridization of probes and target DNA sequences, ligation of the probes, PCR amplification, capillary electrophoresis and data analysis. MLPA has a variety of applications including diagnosis and prognosis of tumors, detection of human genetic diseases, prenatal diagnosis. Due to the increasing occurrence of multiple infections and diseases, there is an urgent need to develop sensitive,specific, and high throughput tools to detect pathogens. MLPA is becoming a promising method in the area of high throughput detection of pathogens. This review gives an overview on the principles and procedures,applications in detection of pathogens and diagnosis of tumors and genetic diseases, advantages and limitations, and perspectives of this technique aiming to provide insights into the application and improvement of MLPA technique in animal pathogen detection and disease diagnosis.

关 键 词：多重连接探针扩增技术(MLPA) 甲基化特异性MLPA(MS-MLPA) 病原体检测 肿瘤诊断 遗传疾病诊断 

分 类 号：S851.3[农业科学—预防兽医学]