短喙型鹅细小病毒感染雏鸭的排毒规律研究

俞博; 林甦; 陈仕龙; 程晓霞; 陈秀琴; 陈少莺

doi:10.19303/j.issn.1008-0384.2018.03.002

短喙型鹅细小病毒感染雏鸭的排毒规律研究

doi: 10.19303/j.issn.1008-0384.2018.03.002

俞博^{1, 2},
林甦^{2, 3},
陈仕龙^{2, 3},
程晓霞^{2, 3},
陈秀琴^{2, 3},
陈少莺^{2, 3, ,}

1.
福建农林大学动物科学学院, 福建福州 250002
2.
福建省农业科学院畜牧兽医研究所, 福建福州 350013
3.
福建省畜禽疫病防治工程技术研究中心, 福建福州 350013

基金项目:

国家重点研发计划 2016YFD0500805-4

福建省科技计划项目——省属公益类科研院所基本科研专项 2017R1023-1

详细信息

作者简介:
俞博(1991-), 男, 硕士研究生, 主要从事动物传染病免疫学研究

通讯作者:
陈少莺(1962-), 女, 研究员, 主要从事动物传染病病原与免疫防控研究(E-mail:chensy58@163.com)

中图分类号: S831.7
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- 被引次数: 0
出版历程
- 收稿日期: 2018-01-19
- 修回日期: 2018-02-11
- 刊出日期: 2018-03-01

Excretion of SBDS-GPV in Infected Ducklings

YU Bo^{1, 2},
LIN Su^{2, 3},
CHEN Shi-long^{2, 3},
CHENG Xiao-xia^{2, 3},
CHEN Xiu-qin^{2, 3},
CHEN Shao-ying^{2, 3
, ,}

1.
College of animal Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
2.
Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture Sciences, Fuzhou, Fujian 350013, China
3.
Fujian Animal Diseases Control Technology Development Center, Fuzhou, Fujian 350013, China

摘要

摘要: 本研究以短喙型鹅细小病毒（SBDS-GPV，M15株）口服感染2日龄健康易感半番鸭，采集感染后（PI）3、5、7、10、12、14、21 d的口咽和泄殖腔棉拭子，应用荧光定量PCR（qPCR）、病毒分离（VI）结合免疫荧光法（IFA）测定雏鸭感染后的排毒情况。结果显示：qPCR法检测SBDS-GPV感染雏鸭的口咽和泄殖腔排毒时间分别为PI 3~10 d和PI 3~14 d；病毒分离结合IFA检测SBDS-GPV感染雏鸭的泄殖腔排毒时间为PI 5~10 d。研究结果在国内外首次明确了雏鸭感染SBDS-GPV后可经口咽和泄殖腔向外界排毒，且泄殖腔排毒水平高于口咽。
- 半番鸭 /
- 短喙矮小综合征(SBDS-GPV) /
- 排毒规律 /
- 荧光定量PCR /
- 病毒分离
Abstract: Two-day-old mule ducklings were orally inoculated with short beak and dwarfism syndrome goose parvovirus (SBDS-GPV, strain M15) for this study. Swab samples from oropharynx and cloaca of the infected birds were collected after 3, 5, 7, 10, 12, 14, and 21 days post infection (dPI) for the virus detection by quantitative real-time PCR (qPCR), virus isolation (VI), and immunofluorescence (IFA). As determined by qPCR, the excretion of SBDS-GPV through oropharynx occurred in 3-10 dPI, and cloaca, in 3-14 dPI. On the other hand, the VI and IFA tests indicated that the cloacal excretion happened in 5-10 dPI. The results, for the first time, demonstrated that SBDS-GPV could pass through both oropharynx and cloaca of a diseased duckling, and that the greater extent of the excretion took place via cloaca.
- mule duckling /
- short beak and dwarfism syndrome goose parvovirus /
- excretion /
- qPCR /
- virus isolation

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图 1 SBDS-GPV致MDEF细胞病变

注：A为Normal MDEF，B为CPE 7 dPI。

Figure 1. CPE of MDEF inoculated with SBDS-GPV

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图 2 SBDS GPV感染MDEF细胞的IFA结果

注：A为Normal MDEF，B为Infected MDEF。

Figure 2. IFA results of MDEF inoculated with SBDS-GPV

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表 1 荧光定量PCR法检测泄殖腔拭子中SBDS-GPV的CT值结果

Table 1. CT values of SBDS-GPV in cloaca swabs detected by qPCR

qPCR	PI 3 d				PI 5 d				PI 7 d				PI 10 d				PI 12 d				PI 14 d				PI 21 d
qPCR	A	B	C	D	A	B	C	D	A	B	C	D	A	B	C	D	A	B	C	D	A	B	C	D	A	B	C	D
CT值	24.17	20.76	22.34	22.59	18.84	19.84	19.12	19.56	25.26	22.19	22.97	24.48	23.57	21.44	22.31	22.70	24.21	26.91	24.45	26.67	26.52	25.24	25.75	26.01	26.68	26.68	26.89	26.47
结果判定	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	-	+	-	-	+	+	-	-	-	-	-
CT均值	22.47				19.34				23.73				22.51					25.56			25.88			26.68
注：“+”表示阳性；“-”表示阴性。下表同。

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表 2 荧光定量PCR法检测口咽拭子中SBDS-GPV的CT值结果

Table 2. CT values of SBDS-GPV in oropharynx swabs detected by qPCR

qPCR	PI 3 d				PI 5 d				PI 7 d				PI 10 d				PI 12 d				PI 14 d				PI 21 d
qPCR	E	F	G	H	E	F	G	H	E	F	G	H	E	F	G	H	A	B	C	D	A	B	C	D	A	B	C	D
CT值	23.30	21.02	22.12	22.2	20.13	20.37	19.98	20.52	24.62	25.81	24.81	25.62	25.07	25.24	25.33	24.98	27.34	29.12	29.01	27.45	28.55	28.97	28.83	28.69	28.12	28.56	28.36	28.32
结果判定	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	-	-	-	-	-	-	-	-	-	-	-	-
CT均值	22.16				20.25				25.22				25.16					28.23			28.76			28.34

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表 3 口咽和泄殖腔棉拭子样品接种MDEF的细胞病变和IFA检测结果

Table 3. CPE and IFA results of MDEF infected with oropharynx and cloaca swab samples

样品	盲传代数	PI 3 d		PI 5 d		PI 7 d		PI 10 d		PI 12 d		PI 14 d		PI 21 d
样品	盲传代数	CPE	IFA	CPE	IFA	CPE	IFA	CPE	IFA	CPE	IFA	CPE	IFA	CPE	IFA
泄殖腔	1代	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	2代	-	-	-	+	-	+	-	+	-	-	-	-	-	-
	3代	-	+	+	+++	+	+++	+	+++	-	-	-	-	-	-
	阳性数	1/4		2/4		3/4		1/4		0/4		0/4		0/4
口咽	1代	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	2代	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	3代	-	-	-	+	-	±	-	-	-	-	-	-	-	-
	阳性数	0/4		1/4		0/4		0/4		0/4		0/4		0/4
注(1)CPE：“+”表示结果为阳性；“-”表示结果为阴性。(2)荧光判定标准：50个视野未见荧光灶(-)；20个视野见1~2荧光灶(±)；10视野见1~2个荧光灶(+)；1个视野见1~2个荧光灶(++)；1个视野见5~10个以上荧光灶(+++)；1个视野见10个以上荧光灶(++++)。

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