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Development of a multiplex RT-PCR assay for the routine detection of seven RNA viruses in Apis mellifera.
Journal of Virological Methods ( IF 2.2 ) Pub Date : 2020-03-20 , DOI: 10.1016/j.jviromet.2020.113858
Abdurrahman Anil Cagirgan 1 , Zafer Yazici 2
Affiliation  

Colony losses in apiaries are frequently one of the most important problems in beekeeping. Colony loss is multifactorial with many reported disorders, Colony Collapse Disorder (CCD), is an increasingly recognised phenomenon which is thought to be caused by many pathogens, including viruses. The aim of this study was to develop a multiplex RT-PCR (mRT-PCR) test to obtain faster results in routine diagnostic laboratories for seven crucial bee viruses. Specific primers for seven RNA viruses, including Israeli acute bee paralysis virus (IAPV), deformed wing virus (DWV), sacbrood virus (SBV), acute bee paralysis virus (ABPV), black queen cell virus (BQCV), kashmir bee virus (KBV) and chronic paralysis virus (CBPV), were used for testing procedure. The mRT-PCR assay can amplify seven plasmid DNA fragments from the pooled viral genomes and it was shown to be sensitive because virus copy numbers were detected to be 104 copies/μl when log10 serial dilutions were performed for the optimized mRT- PCR method. It is concluded that, mRT-PCR test can be used in routine analysis because this assay can perform specific, sensitive and reliable results also achieves economic gains and time due to detecting seven viral agents simultaneously.

中文翻译:

开发用于常规检测蜜蜂蜜蜂中7种RNA病毒的多重RT-PCR分析方法。

养蜂场中的菌落损失通常是养蜂业中最重要的问题之一。菌落丧失是多种因素引起的,有许多报道的疾病,即菌落崩溃紊乱症(CCD),是一种日益被人们认可的现象,被认为是由许多病原体(包括病毒)引起的。这项研究的目的是开发一种多重RT-PCR(mRT-PCR)测试,以便在常规诊断实验室中针对7种重要的蜜蜂病毒获得更快的结果。七种RNA病毒的特异性引物,包括以色列急性蜂麻痹病毒(IAPV),变形翼病毒(DWV),虫病毒(SBV),急性蜂麻痹病毒(ABPV),黑皇后细胞病毒(BQCV),克什米尔蜜蜂病毒(测试程序使用KBV)和慢性麻痹病毒(CBPV)。mRT-PCR分析可从合并的病毒基因组中扩增7个质粒DNA片段,因此显示出敏感性,因为当对优化的mRT-PCR方法进行log10系列稀释时,检测到的病毒拷贝数为104拷贝/μl。结论是,mRT-PCR测试可用于常规分析,因为该检测方法可以执行特异,灵敏和可靠的结果,并且由于同时检测7种病毒制剂,因此可实现经济收益和时间节省。
更新日期:2020-03-20
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