摘要
目的建立蜜蜂黑蜂王台病毒(Black queen cell virus,BQCV)的RT-PCR快速检测方法。方法根据BQCV全基因组序列,在其3′端1 000 bp的区域内设计1对特异性PCR引物,从受BQCV感染的蜜蜂的蛹样品中提取总RNA,进行RT-PCR,并对该法的特异性、敏感性及重复性进行验证。结果从BQCV全基因组序列中可扩增出700 bp的基因条带。该方法可扩增0.1 ng的BQCV DNA;仅能在BQCV中扩增出700 bp的目的基因条带,在蜜蜂急性麻痹病病毒、残翼病病毒、蜜蜂囊状幼虫病病毒中均未扩增出目的基因条带;对相同样品进行多次检测,均在700 bp处出现目的条带。结论已成功建立BQCV RT-PCR快速检测方法,该方法特异性强,敏感性高,重复性好,可用于黑蜂王台病的快速诊断和混合感染的鉴别诊断。
Objective To develop a RT-PCR method for rapid detection of black queen cell virus（BQCV）.Methods According to the whole genome sequence of BQCV,a pair of specific PCR primers were designed in a 1 000 bp region at 3＇-terminus,based on which total RNA was extracted from the bee pupae infected with BQCV for RT-PCR,and the developed method was verified for specificity,sensitivity and reproducibility.Results The gene fragment at a length of 700 bp was amplified from the whole genome of BQCV.A portion of 0.1 ng of BQCV DNA was amplified by the developed method.However,no target gene bands were amplified from acute bee paralysis virus,deformed wing virus or sacbrood virus.The target band at a length of 700 bp was amplified from the same sample repeatedly by the developed method.Conclusion A RT-PCR method for rapid detection of BQCV was successfully developed,which showed high specificity,sensitivity and reproducibility and might be used for rapid diagnosis of BQCV disease and differential diagnosis of mixed infection.
出处
《中国生物制品学杂志》
CAS
CSCD
2011年第10期1227-1229,共3页
Chinese Journal of Biologicals
关键词
黑蜂王台病病毒
逆转录聚合酶链式反应
Black queen cell virus（BQCV）
Reverse transcription-polymerase chain reaction（RT-PCR）
