Correlation of Helicobacter pylori virulence genotypes and clinical characteristics

doi:10.3969/j.issn.1673-8640.2016.06.009

Abstract

Abstract:

Objective To study the molecular characteristics of Helicobacter pylori virulence genotypes and the correlation with different Helicobacter pylori-infected gastroduodenal diseases. Methods A total of 514 gastric mucosa biopsy samples were collected.Helicobacter pylori was isolated. Ten kinds of specific probes for Helicobacter pylori virulence genotypes were designed and synthesized,and the specific virulence genes were detected. The correlation of Helicobacter pylori virulence genotypes and clinical characteristics [chronic superficial gastritis,chronic atrophic gastritis and peptic ulcer disease（PUD）] was analyzed. Results A total of 129 isolates of Helicobacter pylori were isolated from 514 gastric mucosa biopsy samples. Helicobacter pylori carried multiple kinds of virulence genotypes,such as cagA,vacA（m1,m2 and s1）,iceA1,iceA2,dupA,luxS and oipA. Genotype vacA s1m2 and the combined genotype iceA1⁺/iceA2⁺ had an increased risk for PUD. Genotype vacA s1m1 was a risk factor for chronic active gastritis,while dupA was closely associated with duodenal ulcer. Conclusions Helicobacter pylori isolated from Shanghai carrys many kinds of virulence genes,iceA1⁺/iceA2⁺ and vacA s1m2 maybe risk factors for PUD,and dupA could be considered as a duodenal ulcer-specific virulence marker.

Key words: Helicobacter pylori, Virulence genes, Gastroduodenal diseases, Clinical correlation

CLC Number:

R446.5

HU Binjie, ZHAO Fuju, WANG Shiwen, XIANG Ping, YANG Changqing, FANG Yi, CHEN Fei, YANG Feng, ZHAO Hu, ZHANG Yanmei. Correlation of Helicobacter pylori virulence genotypes and clinical characteristics[J]. Laboratory Medicine, 2016, 31(6): 479-485.

Figures/Tables 4

References 29

[1]	PEEK RM Jr, BLASER MJ.Helicobacter pylori and gastrointestinal tract adenocarcinomas[J]. Nat Rev Cancer,2002,2(1):28-37.
[2]	PETERSON WL.Helicobacter pylori and peptic ulcer disease[J]. N Engl J Med,1991,324(15):1043-1048.
[3]	PARSONNET J, HANSEN S, RODRIGUEZ L, et al.Helicobacter pylori infection and gastric lymphoma[J]. N Engl J Med,1994,330(18):1267-1271.
[4]	WONG BC, LAM SK, WONG WM, et al.Helicobacter pylori eradication to prevent gastric cancer in a high-risk region of China: a randomized controlled trial[J]. JAMA,2004,291(2):187-194.
[5]	周丽芳. 16S rRNA等相关基因快速检测幽门螺杆菌及其毒力的多重基因定量分析系统的建立与应用[D]. 上海:复旦大学,2014.
[6]	SHANKS AM, EL-OMAR EM.Helicobacter pylori infection, host genetics and gastric cancer[J]. J Dig Dis,2009,10(3):157-164.
[7]	张万岱,胡伏莲,萧树东,等. 中国自然人群幽门螺杆菌感染的流行病学调查[J]. 现代消化及介入诊疗,2010,15(5):265-270.
[8]	PARSONNET J, FRIEDMAN GD, VANDERSTEEN DP, et al.Helicobacter pylori infection and the risk of gastric carcinoma[J]. N Engl J Med,1991,325(16):1127-1131.
[9]	BLASER MJ.Helicobacter pylori:its role in disease[J]. Clin Infect Dis,1992,15(3):386-391.
[10]	任莉. 宿主体内幽门螺杆菌基因组多态性分析及形成机制研究[D]. 重庆:第三军医大学,2011.
[11]	PROENCA-MODENA JL, ACRANI GO, BROCCHI M.Helicobacter pylori:phenotypes, genotypes and virulence genes[J]. Future Microbiol,2009,4(2):223-240.
[12]	陈莫耶,袁媛. 幽门螺杆菌不同感染阶段的相关毒力因子及其致病性[J]. 世界华人消化杂志,2012(20):2937-2943.
[13]	SHEU BS, YANG HB, YEH YC, et al.Helicobacter pylori colonization of the human gastric epithelium: a bug's first step is a novel target for us[J]. J Gastroenterol Hepatol,2010,25(1):26-32.
[14]	杨瑞宁,操敏,李先富,等. 细菌中LuxS/AI-2密度感应系统分子机制研究进展[J]. 检验医学,2013,28(9):856-861.
[15]	胡玢婕,赵付菊,赵虎. 幽门螺杆菌生物膜形成与其耐药机制的相关性[J]. 检验医学,2014,29(8):865-870.
[16]	成虹,胡伏莲,谢勇,等. 中国幽门螺杆菌耐药状况以及耐药对治疗的影响——全国多中心临床研究[J]. 胃肠病学,2007,12(9):525-530.
[17]	SMITH SM, O'MORAIN C, MCNAMARA D. Antimicrobial susceptibility testing for Helicobacter pylori in times of increasing antibiotic resistance[J]. World J Gastroenterol,2014,20(29):9912-9921.
[18]	张艳梅,胡玢婕,赵付菊,等. 幽门螺杆菌耐药性及其相关耐药基因的研究[J]. 检验医学,2016,31(5):412-418.
[19]	YAMAOKA Y, KODAMA T, GUTIERREZ O, et al.Relationship between Helicobacter pylori iceA, cagA, and vacA status and clinical outcome: studies in four different countries[J]. J Clin Microbiol,1999,37(7):2274-2279.
[20]	SHIOTA S, SUZUKI R, YAMAOKA Y.The significance of virulence factors in Helicobacter pylori[J]. J Dig Dis,2013,14(7):341-349.
[21]	O'TOOLE PW, SNELLING WJ, CANCHAYA C, et al. Comparative genomics and proteomics of Helicobacter mustelae,an ulcerogenic and carcinogenic gastric pathogen[J]. BMC Genomics,2010,11:164.
[22]	YAMAOKA Y.Mechanisms of disease: Helicobacter pylori virulence factors[J]. Nat Rev Gastroenterol Hepatol,2010,7(11):629-641.
[23]	FURUTA Y, KONNO M, OSAKI T, et al.Microevolution of virulence-related genes in Helicobacter pylori familial infection[J]. PLoS One,2015,10(5):e127197.
[24]	ZHENG RN,LI SR,MASAHIRO A.Helicobacter pylori CagA and gastric carcinogenesis[J]. Asian Pac J Cancer Prev,2012,13(12):6305-6310.
[25]	KUSTERS JG, VAN VLIET AH, KUIPERS EJ.Pathogenesis of Helicobacter pylori infection[J]. Clin Microbiol Rev,2006,19(3):449-490.
[26]	ATHERTON JC,CAO P,PEEK RM Jr,et al.Mosaicism in vacuolating cytotoxin alleles of Helicobacter pylori. Association of specific vacA types with cytotoxin production and peptic ulceration[J]. J Biol Chem,1995,270(30):17771-17777.
[27]	LU H,HSU PI,GRAHAM DY,et al.Duodenal ulcer promoting gene of Helicobacter pylori[J]. Gastroenterology,2005,128(4):833-848.
[28]	RADER BA,WREDEN C,HICKS KG,et al.Helicobacter pylori perceives the quorum-sensing molecule AI-2 as a chemorepellent via the chemoreceptor TlpB[J]. Microbiology,2011,157(Pt 9):2445-2455.
[29]	VAN DOORN LJ,FIGUEIREDO C,SANNA R,et al.Clinical relevance of the cagA, vacA, and iceA status of Helicobacter pylori[J]. Gastroenterology,1998,115(1):58-66.

基因		引物序列（5'~3'）	片段大小（bp）
cagA	F:	AATACACCAACGCCTCCAAG	400
cagA	R:	TTGTTGCCGCTTTTGCTCTC	400
vacA s1	F:	ATGGAAATACAACAAACACAC	258
vacA s1	R:	CTGCTTGAATGCGCCAAAC	258
vacA s2	F:	ATGGAAATACAACAAACACAC	286
vacA s2	R:	CTGCTTGAATGCGCCAAAC	286
vacA m1	F:	CAATCTGTCCAATCAAGCGAG	567
vacA m1	R:	GCGTCAAAATAATTCCAAGG	567
vacA m2	F:	CAATCTGTCCAATCAAGCGAG	642
vacA m2	R:	GCGTCAAAATAATTCCAAGG	642
iceA1	F:	GTGTTTTTAACCAAAGTATC	247
iceA1	R:	CTATAGCCASTYTCTTTGCA	247
iceA2	F:	GTTGGGTATATCACAATTTAT	229/334
iceA2	R:	TTRCCCTATTTTCTAGTAGGT	229/334
dupA	F:	ATGAGTTCTATTAACAG	1 839
dupA	R:	TTAAATACTCTTCCTTATAAG	1 839
oipA	F:	CGCGGAAAGGAACGGGTTTT	519
oipA	R:	TTAGCGTCTAGCGTTCTGCC	519
luxS	F:	CTGTGCGCCCTCTAAAGTGT	261
luxS	R:	CACATGGACATGCCTAGCCT	261

基因		引物序列（5'~3'）	片段大小（bp）
cagA	F:	AATACACCAACGCCTCCAAG	400
cagA	R:	TTGTTGCCGCTTTTGCTCTC	400
vacA s1	F:	ATGGAAATACAACAAACACAC	258
vacA s1	R:	CTGCTTGAATGCGCCAAAC	258
vacA s2	F:	ATGGAAATACAACAAACACAC	286
vacA s2	R:	CTGCTTGAATGCGCCAAAC	286
vacA m1	F:	CAATCTGTCCAATCAAGCGAG	567
vacA m1	R:	GCGTCAAAATAATTCCAAGG	567
vacA m2	F:	CAATCTGTCCAATCAAGCGAG	642
vacA m2	R:	GCGTCAAAATAATTCCAAGG	642
iceA1	F:	GTGTTTTTAACCAAAGTATC	247
iceA1	R:	CTATAGCCASTYTCTTTGCA	247
iceA2	F:	GTTGGGTATATCACAATTTAT	229/334
iceA2	R:	TTRCCCTATTTTCTAGTAGGT	229/334
dupA	F:	ATGAGTTCTATTAACAG	1 839
dupA	R:	TTAAATACTCTTCCTTATAAG	1 839
oipA	F:	CGCGGAAAGGAACGGGTTTT	519
oipA	R:	TTAGCGTCTAGCGTTCTGCC	519
luxS	F:	CTGTGCGCCCTCTAAAGTGT	261
luxS	R:	CACATGGACATGCCTAGCCT	261

毒力基因	检出例数	检出率（%）
cagA
cagA⁺	85	98.8
cagA^-	1	1.2
vacA
s1⁺	86	100.0
s2⁺	0	0.0
s1s2	0	0.0
m1⁺	26	30.2
m2⁺	60	69.8
m1m2	0	0.0
s1m1	26	30.2
s1m2	60	69.8
s2m1	0	0.0
s2m2	0	0.0
iceA
iceA1⁺	64	74.4
iceA2⁺	40	46.5
iceA1⁺/ iceA2^-	43	50.0
iceA1^-/ iceA2⁺	19	22.1
iceA1⁺/ iceA2⁺	21	24.4
iceA1^-/ iceA2^-	3	3.5
dupA
dupA⁺	29	33.7
dupA^-	57	66.3
luxS
LuxS⁺	86	100.0
LuxS^-	0	0.0
oipA
oipA⁺	86	100.0
oipA^-	0	0.0

毒力基因	检出例数	检出率（%）
cagA
cagA⁺	85	98.8
cagA^-	1	1.2
vacA
s1⁺	86	100.0
s2⁺	0	0.0
s1s2	0	0.0
m1⁺	26	30.2
m2⁺	60	69.8
m1m2	0	0.0
s1m1	26	30.2
s1m2	60	69.8
s2m1	0	0.0
s2m2	0	0.0
iceA
iceA1⁺	64	74.4
iceA2⁺	40	46.5
iceA1⁺/ iceA2^-	43	50.0
iceA1^-/ iceA2⁺	19	22.1
iceA1⁺/ iceA2⁺	21	24.4
iceA1^-/ iceA2^-	3	3.5
dupA
dupA⁺	29	33.7
dupA^-	57	66.3
luxS
LuxS⁺	86	100.0
LuxS^-	0	0.0
oipA
oipA⁺	86	100.0
oipA^-	0	0.0