15种血清型的鸭疫里默氏杆菌毒力基因检测和耐药性分析

doi:10.11843/j.issn.0366-6964.2026.01.031

Abstract

Abstract:

In order to understand the drug resistance and virulence gene distribution of Riemerella anatipestifer isolates in some areas of China， K-B method was used to determine the sensitivity of 15 serotypes of R. anatipestifer to 10 commonly used antibiotics， and PCR technology was used to detect 16 drug resistance genes and 16 virulence genes of different serotypes of R. anatipestifer isolates. The results showed that the resistance rates of amoxicillin and ampicillin were 93.33% and 80%， respectively. The detection results of drug-resistant genes showed that the detection rates of tet（X） gene of tetracycline， floR gene of amide alcohol and aph（3'）-Ⅱ gene of aminoglycoside were 93.33%， 80% and 73.33% respectively. The results of virulence gene detection showed that tctR， TbdR1， TR， CAMP and AS87-0405 had the highest detection rates， which were 86.67%， 86.67%， 80%， 73.33% and 73.33% respectively. These isolates are sensitive to tetracyclines， quinolones and amide alcohols， and have high resistance to β-lactams. At the same time， these isolates carry a variety of virulence genes， which may indicate that they have strong virulence. The above results provide some reference for the prevention and control of R. anatipestifer in some areas of China.

Key words: Riemerella anatipestifer, virulence gene, drug resistance gene, drug-fast

CLC Number:

S852.61

LI Chenyang, ZHANG Chuanmei, HUI Yutao, LÜ Ying, YANG Min. Detection of Virulence Genes and Drug Resistance Analysis of 15 Serotypes of Riemerella anatipestifer[J]. Acta Veterinaria et Zootechnica Sinica, 2026, 57(1): 360-368.

Figures/Tables 7

Table 1

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Fig.1

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Table 5

Table 6

References 18

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耐药基因 Resistance gene	引物序列（5'→3'） Primer sequences	产物大小/bp Product length	退火温度/℃ Annealing temperature
aac（6′）-Ⅱ	F：TTGCGATGCTCTATGAGTGGCTA	480	52
aac（6′）-Ⅱ	R：CTCGAATGCCTGGCGTGTTT	480	52
aph（3'）-Ⅱa	F：TGACTGGGCACAACAGACA	717	54
aph（3'）-Ⅱa	R：TCAAGAAGGCGATAGAAGGC	717	54
aph（3'）-Ⅱ	F：TCTGAAACATGGCAAAGGTAG	582	54
aph（3'）-Ⅱ	R：AGCCGTTTCTGTAATGAAGGA	582	54
rmtC	F：CGAAGAAGTAACAGCCAAAG	711	55
rmtC	R：ATCCCAACATCTCTCCCACT	711	55
tet （A）	F：GCGCCTTTCCTTTGGGTTCT	831	51
tet （A）	R：CCACCCGTTCCACGTTGTTA	831	51
tet （B）	F：CATTAATAGGCGCATCGCTG	930	55
tet （B）	R：TGAAGGTCATCGATAGCAGG	930	55
tet （C）	F：GCTGTAGGCATAGGCTTGGT	888	55
tet （C）	R：GCCGGAAGCGAGAAGAATCA	888	55
tet （X）	F：ATGACAATGCGAATAGATACAGAC	1 167	53
tet （X）	R：CAATTGCTGAAACGTAAAGTC	1 167	53
blaSHV	F：CACTCAAGGATGTATTGTG	885	55
blaSHV	R：TTAGCGTTGCCAGTGCTCG	885	55
blaDHA	F：AACTTTCACAGGTGTGCTGT	387	56
blaDHA	R：CCGTACGCATACTGGCTTTC	387	56
qnrA	F：ATTTCTCACGCCAGGATTTG	516	58
qnrA	R：GATCGGCAAAGGTTAGGTCA	516	58
qnrB	F：GATCGTGAAAGCCAGAAAGG	469	58
qnrB	R：ACGATGCCTGGTAGTTGTCC	469	58
qnrS	F：ACGACATTCGTCAACTGCAA	417	53
qnrS	R：TAAATTGGCACCCTGTAGGC	417	53
Sul1	F：GTGACGGTGTTCGGCATTCT	779	68
Sul1	R：CCGAGAAGGTGATTGCGCT	779	68
Sul2	F：CCTGTTTCGTCCGACACAGA	435	51
Sul2	R：GAAGCGCAGCCGCAATTCAT	435	51
FloR	F：CTGAGGGTGTCGTCATCTAC	673	58
FloR	R：GCTCCGACAATGCTGACTAT	673	58

目的基因 Target genes	引物序列（5'→3'） Primer sequences	产物大小/bp Product length	退火温度/℃Annealing temperature
luxC	F：CGCGGATCCATGAATATCAGTATTAAAATTTC	1 035	52
luxC	R：CGCGTCGACTTAGTCTATGGTTCTTAGAAAAG	1 035	52
luxr1	F：CGCGGATCCATGGAAGATATCAATGGTTT	759	52
luxr1	R：CGCGTCGACTTAAATCAGTTTATTAATTTTAGC	759	52
AMT	F：CGCGGATCCATGAATATGTTTTTGTAGACG	258	52
AMT	R：CGCGTCGACTCAATCATAATGAAATCTGC	258	52
luxE	F：CGCGGATCCATGCCTTCTATTTTTGATATTA	999	52
luxE	R：CGCGTCGACCTAAGAAACCAAAAGGCTAC	999	52
IS1	F：CGCGGATCCATGATAGAAATTAAGACCTTG	654	52
IS1	R：CGCGTCGACCTAATACCAAAAATATATTTTC	654	52
tctR	F：CGCGGATCCATGAGCAACAGGATATTATT	687	52
tctR	R：CGCGTCGACTTAATTTTTAACTAGAAGCC	687	52
lipidA	F：CGCGGATCCATGCTAAATAATATCCTTTT	885	52
lipidA	R：CGCGTCGACTTTATTGTTTAGCAAAAGTT	885	52
TR	F：CGCGGATCCATGGAAGATATCAATGGTTT	759	52
TR	R：CGCGTCGACTTAAATCAGTTTATTAATTTTAGC	759	52
cwaH	F：CGCGGATCCATGAGAAGATTTATTGGTTT	522	52
cwaH	R：CGCGTCGACTTAAAGTATTCTTTTTGCAA	522	52
CAMP	F：ATGAAACAATCTATTATCTTAGGTA	1 026	55
CAMP	R：TTACTTTGCATTTAACTCATATC	1 026	55
TbdR1	F：TCGTGGGGCAATCTAAACTAAGC	427	55
TbdR1	R：AGCCCTGCCCAAACATCATAA	427	55
wza	F：AGTCTTTAGACAAGTAGATGCC	936	50
wza	R：TTACTTTGCATTTAACTCATATC	936	50
AS87-0405	F：AGGCCGCTTTAGGTTCTGTTC	585	55
AS87-0405	R：CTTCGCCTATTTTCTCATCGTATTT	585	55
SIP	F：ATGCCTAAGACACCGAAATGGATG	729	55
SIP	R：TTACAAACCTTGTTTTCCTTCCACC	729	55
luxr2	F：CGCGGATCCATGGGGGATTTTGACGGT	759	52
luxr2	R：CGCGTCGACTTACACTAATTTATTAATTTTAG	759	52
Fur	F：ATGGAACATCAAGAGAAAGATATAG	468	49
Fur	R：TTATGCTTTTTTATGACCGTAGA	468	49

序号 No.	地区 District	分离时间 Isolation time	家禽品种 Breed	日龄/d Age （Day）	血清型 Serotype
1	福建	2022-09	肉鸭	282	14
2	山东	2022-10	肉鸭	34	6
3	山东	2022-10	肉鸭	29	11
4	山东	2022-10	肉鸭	33	13
5	河北	2022-10	肉鸭	27	12
6	河北	2022-11	肉鸭	32	10
7	广东	2022-11	肉种鸡	56	4
8	广西	2022-12	肉鸭	32	16
9	广东	2023-01	鹅	18	1
10	山东	2023-02	肉鸭	24	15
11	广西	2023-12	肉鸭	20	5
12	山东	2022-09	肉鸭	27	17
13	河北	2023-08	蛋鸡	250	7
14	山东	2023-05	肉鸭	24	9
15	福建	2023-12	肉鸭	18	2

药物名称 Drug name	血清型Serotype															耐药率/% Drug resistance rate
药物名称 Drug name	1	2	4	5	6	7	9	10	11	12	13	14	15	16	17	耐药率/% Drug resistance rate
链霉素 Streptomycin	S	R	R	S	R	S	S	S	S	S	S	R	S	S	S	26.67
庆大霉素 Gentamicin	S	S	R	S	S	S	S	S	S	S	S	S	R	S	S	13.33
多西环素 Doxycycline	S	S	I	R	R	S	S	S	I	S	S	S	S	S	S	13.33
替加环素 Tigecycline	S	S	S	S	S	S	S	S	S	S	S	S	S	S	S	0
氨苄西林 Ampicillin	R	R	R	S	R	R	R	I	R	R	R	R	R	R	S	80
阿莫西林 Amoxicillin	R	R	R	R	R	R	R	R	R	R	R	R	I	R	R	93.33
恩诺沙星 Enrofloxacin	S	S	S	S	S	I	S	S	I	S	R	S	R	S	I	13.33
环丙沙星 Ciprofloxacin	S	S	S	S	S	I	R	S	I	S	I	S	I	S	S	6.67
复方新诺明 Co-trimoxazole	S	R	R	S	R	R	R	R	S	R	S	R	R	R	R	73.33
氯霉素 Chloramphenicol	S	I	I	S	S	S	S	S	S	I	S	S	R	S	S	6.67

毒力基因 Virulence genes	血清型 Serotype															检出率/% Detectionrate
毒力基因 Virulence genes	1	2	4	5	6	7	9	10	11	12	13	14	15	16	17	检出率/% Detectionrate
luxC															+	6.67（1/15）
luxr					+							+			+	20.00（3/15）
AMT					+			+			+	+				26.67（4/15）
luxE	+				+	+	+	+	+	+	+					53.33（8/15）
ISl					+		+		+	+	+	+		+		46.67（7/15）
tctR	+	+		+	+	+	+	+	+	+	+	+		+	+	86.67（13/15）
lipidA							+	+	+	+	+	+		+		46.67（7/15）
TR	+	+			+	+		+	+	+	+	+		+	+	73.33（11/15）
cwaH	+					+		+	+		+	+			+	46.67（7/15）
CAMP	+	+	+	+	+	+	+	+		+	+	+		+		80（12/15）
TbdR1		+	+		+	+	+	+	+	+	+	+	+	+	+	86.67（13/15）
wza				+	+			+			+				+	33.33（5/15）
AS87-0405		+	+	+			+	+		+		+		+		53.33（8/15）
SIP	+		+		+	+	+	+	+		+	+		+	+	73.33（11/15）
luxr2			+					+			+					20.00（3/15）
Fur					+	+	+	+	+		+	+				46.67（7/15）
合计Total	6	5	5	4	11	8	9	13	9	8	13	12	1	8	8

Detection of Virulence Genes and Drug Resistance Analysis of 15 Serotypes of Riemerella anatipestifer

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耐药基因 Resistance gene	血清型Serotype															检出率/% Detection rate
耐药基因 Resistance gene	1	2	4	5	6	7	9	10	11	12	13	14	15	16	17	检出率/% Detection rate
aac（6'）-Ⅱ			+													6.67
aph（3'）-ILa
aph（3'）-Ⅱ	+			+	+	+	+	+	+		+	+		+	+	73.33
rmtC
tet （A）
tet （B）
tet （C）												+				6.67
tet（X）		+	+	+	+	+	+	+	+	+	+	+	+	+	+	93.33
blaSHV
blaDHA
qnrA
qnrB											+					6.67
qnrS
Sul1			+													6.67
Sul2			+													6.67
floR	+	+				+	+	+	+	+	+	+	+	+	+	80
合计 Total	2	2	4	2	2	3	3	3	3	2	4	4	2	3	3

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