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浙江大学学报（农业与生命科学版）
Journal of Zhejiang University (Agriculture and Life Sciences)  2019, Vol. 45 Issue (3): 385-390    DOI: 10.3785/j.issn.1008-9209.2018.07.181
Animal sciences & veterinary medicine     
Establishment and application of a polymerase chain reaction method for detection of Melissococcus pluton in honeybee (Apis cerana cerana) in Chun’an County of Zhejiang Province
Rui GUO1(),Jie DONG1,Haikun GUO2,Junzhu LI3,Deqian WANG1()
1. Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. Institute of Quality and Standard for Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Hao Qing-mi Beekeeping Professional Cooperative of Qiandao Lake of Zhejiang, Hangzhou 311701, China
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Abstract  

A polymerase chain reaction (PCR) method was developed to detect Melissococcus pluton and an epidemiological investigation of European foulbrood (EFB) was done in the main bee (Apis cerana cerana) breeding area of Chun’an County, Zhejiang Province. Results indicated that the newly developed method was specific, sensitive, repeatable and accuracy. The epidemiological survey results showed that a total of 11 townships were infected by M. pluton among the 298 bee samples in 17 townships of Chun’an County. The total infection rate was 25.2%, of which EFB infection rate of five townships was ≥40%. In a word, the newly established method for PCR detection makes the infection status of EFB clear in Chun’an County, and it lays a scientific basis for formulating rationally prevention measures of EFB.

Key words： Apis cerana cerana      Melissococcus pluton      European foulbrood      polymerase chain reaction      Chun’an County      epidemiology     
Received: 18 July 2018      Published: 25 June 2019
CLC:  S 895.132  
Corresponding Authors: Deqian WANG     E-mail: 754894816@qq.com;wangdeqian2002@sohu.com
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Rui GUO
Jie DONG
Haikun GUO
Junzhu LI
Deqian WANG
Cite this article:

Rui GUO,Jie DONG,Haikun GUO,Junzhu LI,Deqian WANG. Establishment and application of a polymerase chain reaction method for detection of Melissococcus pluton in honeybee (Apis cerana cerana) in Chun’an County of Zhejiang Province. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(3): 385-390.

URL:

http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2018.07.181     OR     http://www.zjujournals.com/agr/Y2019/V45/I3/385


浙江省淳安县中华蜜蜂蜂房球菌聚合酶链式反应检测方法的建立与应用

通过建立蜂房球菌（Melissococcus pluton）的聚合酶链式反应（polymerase chain reaction, PCR）检测方法，对浙江省淳安县主要中华蜜蜂（Apis cerana cerana）饲养区进行欧洲幼虫腐臭病的流行病学调查。结果显示，新建立的蜂房球菌PCR检测方法特异性强，灵敏度高，并具有良好的重复性和准确性。流行病学调查表明，在淳安县17个乡镇的298群蜜蜂样品中，共有11个乡镇检出蜂房球菌，检出率为25.2%，其中5个乡镇的阳性检出率≥40%。总之，本研究建立了一个新的蜂房球菌PCR检测方法，明确了淳安县中华蜜蜂对欧洲幼虫腐臭病的感染情况，为合理制定该病害的防治措施提供了科学依据。


关键词： 中华蜜蜂,  蜂房球菌,  欧洲幼虫腐臭病,  聚合酶链式反应,  淳安县,  流行病学 
Fig. 1 Optimization of annealing temperatures for PCR
Fig. 2 Identification result of recombinant plasmid of pMD19-T/M. pluton by double restriction-enzyme digestion
Fig. 3 Results of sensitivity test
Fig. 4 Specificity test result of PCR
Fig. 5 Test results of some samples of M. pluton by PCR

序号

No.

乡镇

Town

蜂群（群）

Colony

阳性数（群）

Positive number

蜂房球菌检出率

Positive rate/%
合计 Total 298 75 25.2
1 威坪镇 10 4 40.0
2 千岛湖镇 30 8 26.7
3 金峰乡 30 18 60.0
4 临岐镇 30 15 50.0
5 宋村乡 20 8 40.0
6 文昌镇 10 0 0
7 安阳乡 10 4 40.0
8 里商乡 15 0 0
9 鸠坑乡 18 0 0
10 富文乡 15 3 20.0
11 浪川乡 10 0 0
12 姜家镇 20 0 0
13 左口乡 15 0 0
14 瑶山乡 10 3 30.0
15 屏门乡 10 2 20.0
16 枫树岭镇 30 6 20.0
17 界首乡 15 4 26.7
Table 1 Information of collected samples and detection result of M. pluton
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