论文部分内容阅读
目的：对法国赠送的戊型肝炎病毒（ｈｅｐａｔｉｔｉｓＥｖｉｒｕｓ，ＨＥＶ）Ｆ８６株进行生物学性状与基因特征的鉴定，并与我国分离的８７Ａ株病毒进行比较。方法：用常量法测定病毒的血凝滴度；微量滴定法测定在２ＢＳ、Ａ５４９、ＬＬＣ－ＭＫ２细胞中的病毒滴度；腹腔注射ＢＡＬＢ／ｃ小鼠，３周后取血清进行抗体检测；利用ＲＴ－ＰＣＲ方法从接种病毒的细胞培养液中直接检测病毒ＲＮＡ，ＰＣＲ阳性产物纯化回收后克隆并测序。结果：Ｆ８６株病毒血凝试验阴性；在２ＢＳ、Ａ５４９细胞中连传３代，病毒滴度稳定，在ＬＬＣ－ＭＫ２细胞中病毒滴度稍低；在接种Ｆ８６株病毒的ＢＡＬＢ／ｃ小鼠中可检出特异性抗体；在其细胞培养物中检出ＨＥＶ聚合酶区一段核苷酸序列，测序结果与我国８７Ａ株的同源性为９８．７％。结论：Ｆ８６株病毒不能凝集人Ｏ型红细胞，对２ＢＳ、Ａ５４９及ＬＬＣ－ＭＫ２细胞均敏感，对ＢＡＬＢ／ｃ小鼠不致病。部分序列的测定结果表明该株病毒确为ＨＥＶ，从而由国外实验室证实ＨＥＶ可用人源性细胞分离。
OBJECTIVE: To identify the biological traits and genetic characteristics of hepatitis E virus (HEV) F86 presented in France and to compare with 87A isolates isolated from China. Methods: The titer of virus was determined by the constant method. The titer of virus in 2BS, A549 and LLC-MK2 cells was determined by microtiter method. The BALB / c mice were injected intraperitoneally with the serum for antibody detection after 3 weeks. The viral RNA was directly detected by RT-PCR from the virus-inoculated cell culture medium. The PCR positive product was purified and recovered, then cloned and sequenced. Results: The hemagglutination test of F86 strain was negative. In 2BS, A549 cells were passaged for 3 passages, the virus titer was stable and the titer of virus in LLC-MK2 cells was slightly lower. In BALB / c mice vaccinated with F86 virus Specific antibody could be detected. A nucleotide sequence of HEV polymerase was detected in its cell culture. The sequencing result was 98.7% homologous to 87A in China. CONCLUSION: F86 strain can not agglutinate human O-type erythrocytes and is sensitive to 2BS, A549 and LLC-MK2 cells, but not to BALB / c mice. The results of partial sequence analysis showed that the strain of virus was indeed HEV, which was confirmed by foreign laboratories for the isolation of HEV-available human cells.