Li Weihong, Cao Shouchun, Zhang Quanfu, Li Jiandong, Zhang Shuo, Wu Wei, Qu Jing, Li Chuan, Liang Mifang, Li Dexin
Key Laboratory for Medical Virology, National Health and Family Planning Commission of the People's Republic of China; Laboratory for Viral Hemorrhagic Fever, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, 102206, People's Republic of China.
Institute for Infectious Disease and Endemic Disease Control, Beijing CDC, Beijing, 100013, People's Republic of China.
Virol J. 2017 Jul 18;14(1):133. doi: 10.1186/s12985-017-0799-0.
Hantaan and Seoul viruses, in the Hantavirus genus, are known to cause hemorrhagic fever with renal syndrome (HFRS). The plaque reduction neutralization test (PRNT), as conventional neutralization test for hantaviruses, is laborious and time-consuming. Alternatives to PRNT for hantaviruses are required.
In this study, the methods for Hantaan and Seoul viruses serological typing including microneutralization test (MNT), pseudoparticle neutralization test (PPNT) and immunofluorescence assay based on viral glycoproteins (IFA-GP) were developed and compared with PRNT using a panel of 74 sera including 44 convalescent sera of laboratory confirmed HFRS patients and 30 patients sera of non-hantavirus infection. Antibody titres and serotyping obtained with different methods above were analyzed by paired-t, linear correlation, McNemar χ and Kappa agreement tests.
Antibody titres obtained with MNT, PPNT and IFA-GP were significantly correlated with that obtained with PRNT (p < 0.001). GMT determined by PPNT was statistically higher than that determined by PRNT (p < 0.001), while GMT determined by MNT and IFA-GP were equal with (p > 0.05) and less than (p < 0.001) that obtained with PRNT respectively. Serotyping obtained with MNT and PRNT, PPNT and PRNT were highly consistent (p < 0.001), whereas that obtained with IFA-GP and PRNT were moderately consistent (p < 0.001). There were no significant differences for serotyping between PRNT and MNT, as well as PRNT and PPNT (p > 0.05). IFA-GP was less sensitive than PRNT and MNT for serotyping of hantaviruses infection (p < 0.05). However, for 79.5% (35/44) samples, serotyping determined by IFA-GP and PRNT were consistent.
MNT and PPNT both can be used as simple and rapid alternatives to PRNT, and MNT is more specific while PPNT is more sensitive than other assays for neutralizing antibody determination. So far, this work has been the most comprehensive comparison of alternatives to PRNT.
汉坦病毒属中的汉滩病毒和汉城病毒可引起肾综合征出血热（HFRS）。空斑减少中和试验（PRNT）作为汉坦病毒的传统中和试验，费力且耗时。需要有PRNT的替代方法来检测汉坦病毒。
在本研究中，开发了包括微量中和试验（MNT）、假病毒颗粒中和试验（PPNT）和基于病毒糖蛋白的免疫荧光试验（IFA-GP）在内的汉滩病毒和汉城病毒血清学分型方法，并与PRNT进行比较，使用了一组74份血清，其中包括44份实验室确诊的HFRS患者的恢复期血清和30份非汉坦病毒感染患者的血清。通过配对t检验、线性相关分析、McNemar χ检验和Kappa一致性检验分析上述不同方法获得的抗体滴度和血清学分型结果。
MNT、PPNT和IFA-GP获得的抗体滴度与PRNT获得的抗体滴度显著相关（p < 0.001）。PPNT测定的几何平均滴度（GMT）在统计学上高于PRNT测定的GMT（p < 0.001），而MNT和IFA-GP测定的GMT分别与PRNT相等（p > 0.05）和低于PRNT（p < 0.001）。MNT与PRNT、PPNT与PRNT获得的血清学分型高度一致（p < 0.001），而IFA-GP与PRNT获得的血清学分型中度一致（p < 0.001）。PRNT与MNT以及PRNT与PPNT之间的血清学分型无显著差异（p > 0.05）。IFA-GP在汉坦病毒感染血清学分型方面比PRNT和MNT敏感性低（p < 0.05）。然而，对于79.5%（35/44）的样本，IFA-GP和PRNT确定的血清学分型一致。
MNT和PPNT均可作为PRNT的简单快速替代方法，MNT在中和抗体测定方面更具特异性，而PPNT比其他检测方法更敏感。到目前为止，这项工作是对PRNT替代方法最全面的比较。
