Chanyasanha C, Hasebe F, Matias R, Igarashi A
Department of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, Thailand.
Acta Virol. 1999 Aug;43(4):213-8.
Viral antigen production was examined in the culture fluid of Aedes albopictus clone C6/36 cell line incubated at 28 degrees C and 37 degrees C after infection with four strains of dengue type 4 (DEN-4) virus which were isolated from patients with different clinical severities. During the observation period from day 1 to day 18, the number of infected cells at each day for all four strains did not show any significant difference (P >0.05). Antigen production as determined by the hemagglutination (HA) test and sandwich enzyme linked immunosorbent assay (ELISA) was higher at 28 degrees C than at 37 degrees C for all four DEN-4 virus strains examined. The amount of viral antigen produced was highest for CT93-74 strain (dengue hemorrhagic fever syndrome (DHF) grade II) and was significantly different in comparison to other strains (P <0.001). This strain was followed by CT93-158 and CT93-129 strains (both of DHF grade I), and CT93-77 strain (dengue fever (DF)). The viral antigen production was apparently proportional to the clinical severity of the patient from whom the virus was isolated. These results show that CT93-74 strain could be used to produce DEN-4 virus antigen of sufficiently high titer in the C6/36 cell culture instead of classical extraction of this antigen from suckling mouse brains.
用从不同临床严重程度的登革热4型(DEN - 4)病毒患者中分离出的4株病毒感染白纹伊蚊克隆C6/36细胞系,并在28℃和37℃下培养,检测培养液中的病毒抗原产生情况。在第1天至第18天的观察期内,所有4株病毒每天的感染细胞数均无显著差异(P>0.05)。通过血凝试验(HA)和夹心酶联免疫吸附测定(ELISA)测定,对于所有检测的4株DEN - 4病毒株,28℃时的抗原产生量高于37℃。CT93 - 74株(登革出血热综合征(DHF)II级)产生的病毒抗原量最高,与其他毒株相比有显著差异(P<0.001)。该毒株之后依次是CT93 - 158和CT93 - 129株(均为DHF I级)以及CT93 - 77株(登革热(DF))。病毒抗原产生量显然与分离出病毒的患者的临床严重程度成正比。这些结果表明,CT93 - 74株可用于在C6/36细胞培养中产生足够高滴度的DEN - 4病毒抗原,而无需从乳鼠脑中经典提取该抗原。