福建省牡蛎疱疹病毒(OsHV-1)的检测与分析

doi:10.14012/j.cnki.fjsc.2023.04.008

摘要/Abstract

摘要：

随着牡蛎养殖产业规模的不断扩大,牡蛎养殖病害问题日益严重。近年来,牡蛎疱疹病毒1型(Ostreid herpesvirus 1,OsHV-1)在牡蛎中的感染事件频繁发生,特别是在幼苗期,给牡蛎养殖者造成了经济损失。本文通过普通PCR和实时荧光定量PCR方法对福建省具有一定代表性的6个育苗场及3个主要牡蛎养殖海区的长牡蛎(又称太平洋牡蛎,Crassostrea gigas)和福建牡蛎(又称葡萄牙牡蛎,C.angulata)共90个样本开展OsHV-1检测。结果显示,牡蛎中OsHV-1的平均检出率约11.11%,其中幼苗样本30个,阳性检出率为23.33%,高于成贝中5%的阳性检出率。幼苗阳性样本的病毒含量明显高于成贝,其中牡蛎幼苗样本中OsHV-1最高含量达6.1×10⁷copies/mg。本研究可为牡蛎疱疹病毒的分子学检测及牡蛎疫病防控方面提供参考。

关键词: 牡蛎, 牡蛎疱疹病毒, PCR

Abstract:

With the continuous expansion of oyster culture industry, the problem of oyster disease is becoming more and more serious. In recent years, ostreid herpesvirus 1 (OsHV-1) caused mass death of shellfish, which brought great economic losses to shellfish aquaculture industry. OsHV-1 infection occurred frequently in oysters, especially in the seedling stage. In this paper, 90 samples of Crassostrea gigas and C.angulata from six seedling growing enterprises and three sea areas in Fujian were tested for OsHV-1 by PCR and quantitative real-time PCR. The results showed that the average detection rate of OsHV-1 was about 11.11%. Among them, there were 30 seedling samples, while the positive detection rate of seedling samples was 23.33%, which was higher than the positive detection rate of 5% in adult shellfish, and the infection amount of positive samples in seedling was significantly higher than that in adult shellfish. The infection amount of OsHV-1 in C. gigas seedling sample was the highest, reached 6.1×10⁷copies/mg. This study could provide reference for the detection of ostreid herpesvirus as well as the prevention and control of oyster diseases.

Key words: oyster, ostreid herpesvirus 1(OsHV-1), PCR

中图分类号:

S944.4

贾圆圆. 福建省牡蛎疱疹病毒(OsHV-1)的检测与分析[J]. 渔业研究, 2023, 45(4): 378-384.

JIA Yuanyuan. Detection and analysis of ostreid herpesvirus 1 (OsHV-1) in Fujian Province[J]. Journal of Fisheries Research, 2023, 45(4): 378-384.

图/表 7

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采集地点 Collection site		牡蛎品种 Oyster variety	样本数 Quantity	样品编号 Sample number
育苗场 Hatchery	漳浦(A)	长牡蛎(苗期)	5	1~5
	漳浦(B)	长牡蛎(苗期)	5	6~10
	漳浦(C)	福建牡蛎(苗期)	5	11~15
	诏安(D)	福建牡蛎(苗期)	5	16~20
	莆田(E)	福建牡蛎(苗期)	5	21~25
	莆田(F)	福建牡蛎(苗期)	5	26~30
海区 Sea area	平潭	长牡蛎(成贝)	20	31~50
	深沪湾	福建牡蛎(成贝)	20	51~70
	漳浦	福建牡蛎(成贝)	20	71~90

采集地点 Collection site		牡蛎品种 Oyster variety	样本数 Quantity	样品编号 Sample number
育苗场 Hatchery	漳浦(A)	长牡蛎(苗期)	5	1~5
	漳浦(B)	长牡蛎(苗期)	5	6~10
	漳浦(C)	福建牡蛎(苗期)	5	11~15
	诏安(D)	福建牡蛎(苗期)	5	16~20
	莆田(E)	福建牡蛎(苗期)	5	21~25
	莆田(F)	福建牡蛎(苗期)	5	26~30
海区 Sea area	平潭	长牡蛎(成贝)	20	31~50
	深沪湾	福建牡蛎(成贝)	20	51~70
	漳浦	福建牡蛎(成贝)	20	71~90

引物名称 Primer name	序列(5’-3’) Sequence(5’-3’)
C2	CTCTTTACCATGAAGATACCCACC
C6	GTGCACGGCTTACCATTTTT
BF	GTCGCATCTTTGGATTTAACAA
B4	ACTGGGATCCGACTGACAAC
B-P	6FAM-TGCCCCTGTCATCTTGAGGTATAGACAATC-BHQ-1

引物名称 Primer name	序列(5’-3’) Sequence(5’-3’)
C2	CTCTTTACCATGAAGATACCCACC
C6	GTGCACGGCTTACCATTTTT
BF	GTCGCATCTTTGGATTTAACAA
B4	ACTGGGATCCGACTGACAAC
B-P	6FAM-TGCCCCTGTCATCTTGAGGTATAGACAATC-BHQ-1

成分 Composition	体积/μL Volume
2× PCR Mix	12.5
Upstream primer	1.0
下游引物/10 μmol/L Downstream primer	1.0
无菌双蒸水 Sterile double steaming water	9.5
模板 Template	1.0
总计 Total	25.0