中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2011年
3期
234-240
,共7页
范张洁%段广才%张卫东%郗园林
範張潔%段廣纔%張衛東%郗園林
범장길%단엄재%장위동%치완림
肠道病毒71型%基因型%VP1基因%2A基因%遗传进化
腸道病毒71型%基因型%VP1基因%2A基因%遺傳進化
장도병독71형%기인형%VP1기인%2A기인%유전진화
Enterovirus type 71(EV71)%Genotype%VP1 genes%2 A genes%Phylogenetic
目的 检测临床分离的肠道病毒71型(EV71)轻型、重型毒株VP1基因、2A基因的核苷酸序列,进行遗传进化途径分析.方法 将50份临床手足口病患者粪便标本处理后分离病毒,用EV71特异性引物进行RT-PCR扩增,获得EV71毒株,针对不同临床背景的标本,分别选取轻型、重型的毒株,用特异性引物分别对VP1基因及2A基因进行RT-PCR扩增,对其产物进行测序及遗传进化分析,并探讨它们之间的差别.结果 50份临床手足口病标本中有30份检测是EV71,其中轻型(手足口)13份,重型(手足口并发脑炎或心肌炎)17份.轻型和重型中各自选取5株对VP1基因和2A基因进行测序和遗传学分析,通过同源性比较和构建系统发生树发现,此10株EV71病毒和中国大陆已发表的5株EV71病毒(fuyangEU703814.1、xi_anHM003207.1、shandongEU753418.1、shenzhen-FJ607337.1、henanGU366191.1)全部属于C基因型,且核苷酸同源性较高,VP1基因和2A基因的同源性分别在94.7%~99.4%和93.6%~99.3%范围内.本次分离的10株EV71病毒与A、B基因型代表株比较,核苷酸同源性分别为81.0%~84.6%和78.4%~82.2%,差异较大.与已知的C1、C2、C3亚型代表株比较,核苷酸同源性在87.8%~90.2%,差异≥10%,与已知的C4亚型代表株比较,核苷酸同源性在96.8%~99.6%,因此认为可将这10株病毒划分为C4亚型.在系统发生树上,这10株病毒形成一个较独立的分支.结论 EV71 C4亚型病毒在中国大陆有较广泛的传播,且毒株之间VP1基因的遗传关系紧密;2A基因在轻型和重型病例毒株之间遗传差异不大.
目的 檢測臨床分離的腸道病毒71型(EV71)輕型、重型毒株VP1基因、2A基因的覈苷痠序列,進行遺傳進化途徑分析.方法 將50份臨床手足口病患者糞便標本處理後分離病毒,用EV71特異性引物進行RT-PCR擴增,穫得EV71毒株,針對不同臨床揹景的標本,分彆選取輕型、重型的毒株,用特異性引物分彆對VP1基因及2A基因進行RT-PCR擴增,對其產物進行測序及遺傳進化分析,併探討它們之間的差彆.結果 50份臨床手足口病標本中有30份檢測是EV71,其中輕型(手足口)13份,重型(手足口併髮腦炎或心肌炎)17份.輕型和重型中各自選取5株對VP1基因和2A基因進行測序和遺傳學分析,通過同源性比較和構建繫統髮生樹髮現,此10株EV71病毒和中國大陸已髮錶的5株EV71病毒(fuyangEU703814.1、xi_anHM003207.1、shandongEU753418.1、shenzhen-FJ607337.1、henanGU366191.1)全部屬于C基因型,且覈苷痠同源性較高,VP1基因和2A基因的同源性分彆在94.7%~99.4%和93.6%~99.3%範圍內.本次分離的10株EV71病毒與A、B基因型代錶株比較,覈苷痠同源性分彆為81.0%~84.6%和78.4%~82.2%,差異較大.與已知的C1、C2、C3亞型代錶株比較,覈苷痠同源性在87.8%~90.2%,差異≥10%,與已知的C4亞型代錶株比較,覈苷痠同源性在96.8%~99.6%,因此認為可將這10株病毒劃分為C4亞型.在繫統髮生樹上,這10株病毒形成一箇較獨立的分支.結論 EV71 C4亞型病毒在中國大陸有較廣汎的傳播,且毒株之間VP1基因的遺傳關繫緊密;2A基因在輕型和重型病例毒株之間遺傳差異不大.
목적 검측림상분리적장도병독71형(EV71)경형、중형독주VP1기인、2A기인적핵감산서렬,진행유전진화도경분석.방법 장50빈림상수족구병환자분편표본처리후분리병독,용EV71특이성인물진행RT-PCR확증,획득EV71독주,침대불동림상배경적표본,분별선취경형、중형적독주,용특이성인물분별대VP1기인급2A기인진행RT-PCR확증,대기산물진행측서급유전진화분석,병탐토타문지간적차별.결과 50빈림상수족구병표본중유30빈검측시EV71,기중경형(수족구)13빈,중형(수족구병발뇌염혹심기염)17빈.경형화중형중각자선취5주대VP1기인화2A기인진행측서화유전학분석,통과동원성비교화구건계통발생수발현,차10주EV71병독화중국대륙이발표적5주EV71병독(fuyangEU703814.1、xi_anHM003207.1、shandongEU753418.1、shenzhen-FJ607337.1、henanGU366191.1)전부속우C기인형,차핵감산동원성교고,VP1기인화2A기인적동원성분별재94.7%~99.4%화93.6%~99.3%범위내.본차분리적10주EV71병독여A、B기인형대표주비교,핵감산동원성분별위81.0%~84.6%화78.4%~82.2%,차이교대.여이지적C1、C2、C3아형대표주비교,핵감산동원성재87.8%~90.2%,차이≥10%,여이지적C4아형대표주비교,핵감산동원성재96.8%~99.6%,인차인위가장저10주병독화분위C4아형.재계통발생수상,저10주병독형성일개교독립적분지.결론 EV71 C4아형병독재중국대륙유교엄범적전파,차독주지간VP1기인적유전관계긴밀;2A기인재경형화중형병례독주지간유전차이불대.
Objective To detect VP1 and 2A genes of Enterovirus type 71 (EV71) isolated from clinical specimens of patients with light or heavy symptoms and analyze the homogeneity and phylogenetic tree. Methods Fifty clinical specimens of children with hand-foot-and-mouth disease ( HFMD) were dealed with, which were tested by RT-PCR assay with specific primer pairs for EV71. EV71 isolates from patients with light or heavy clinical symptoms were tested by RT-PCR assay with two specific primer pairs for VP1 and 2A genes of EV71 respectively. All of the PCR products were sequenced and compared with that of previously isolated EV71 isolates available from GenBank by homogeneity and phylogenetic tree analyses. Results The RT-PCR results indicated that 30 isolates were EV71, 13 of 30 isolates were from clinical specimens of patients with light symptoms of hand-foot and mouth, the other were from clinical specimens of patients with heavy symptoms of complications. VP1 genes and 2A genes of 10 EV71 isolated strains including 5 light strains and 5 heavy strains were sequenced and compared with that of previously isolated 5 EV71 Chinese isolates available from GenBank (fuyangEU703814.1, xi_anHM003207. 1, shandongEU753418.1, shenzhenFJ607337.1, henanGU366191. 1) by homogeneity and phylogenetic tree analyses. The homogeneity of VP1 and 2A genes of the 10 EV71 isolated strains and 5 previously isolated strains were between 94.7% -99.4% and 93.6% -99.3% respectively, with the representative isolates of A and B genotypes was between 81.0%-84. 6% and 78. 4%-82. 2% respectively. The data suggested that all of the 10 Chinese isolates belong to EV71 genotype C. There were only 87.8% -90.2% homology among these 10 strains and the representative strains of C1, C2, C3 sub-genotypes of EV71 but 96. 8% -99.6% homology among these 10 strains and the representative strains of C4 sub-genotypes of EV71, this suggested that these 10 Chinese isolates composed the C4 sub-genotype, of the C genotype, that formed a single branch in the phylogenetic tree. Conclusion EV71 of sub-genotype C4 distributed in Mainland China, and VP1 genes have close genetic relationship between isolated strains. There is no obvious difference in 2A genes between clinical specimens of patients with light or heavy symptoms by homogeneity and phylogenetic tree analyses.