Establishment of indirect immunofluorescence assay (IFA) for detection of IgG antibody against new bunyavirus
Huang, X.-Y.; Du, Y.-H.; Li, X.-l.; Ma, H.; Man, R.-Q.; Kang, K.; Tang, X.-Y.; Chen, H.-M.; Liu, G.-H.; Xu, B.-L.
Zhonghua Yu Fang Yi Xue Za Zhi 46(2): 165-168
2012
ISSN/ISBN: 0253-9624 PMID: 22490201 Document Number: 657919
To develop an indirect immunofluorescence assay (IFA) for detection of IgG antibodies against new bunyavirus. The antigen slides were prepared with 5 new bunyavirus strains isolated using Africa green monkey kidney (Vero) cells. Specificity and sensitivity evaluation of IFA were carried out by optimizing working conditions of IFA. Using established IFA, serum samples from both acute and recovery phases were tested for 126 cases with fever thrombocytopenia and leukopenia syndrome in Xinyang, Henan province in 2007 - 2011. The results were compared with detections by RT-PCR. The new bunyavirus stable immunofluorescence specific WZ69 strain was selected to prepare antigen slides of IFA. The optimum conditions of IFA were: optimum dilution for primary antibody (serum) and secondary antibody (isosulfocyanic acid fluorescence marked goat anti-human IgG antibody) was 1:40 and 1:150 respectively. The optimum dilution for Evans blue in secondary antibody was 1:20 000. Among the 126 patients, 96 paired serum specimens were tested positive to the new bunyavirus and 30 patients were tested negative to the virus. The positive rate of antibodies was 76.19%. There was no significant difference in results between IFA and RT-PCR (72.22% (91/126)) (P > 0.05). The IFA has high sensitivity and specificity with easy operation. It can be used in detecting the new bunyavirus infection in patients with fever, thrombocytopenia and leukopenia syndrome.