Establishment of indirect immunofluorescence assay (IFA) for detection of IgG antibody against new bunyavirus.
- Author:
Xue-Yong HUANG
1
;
Yan-Hua DU
;
Xing-le LI
;
Hong MA
;
Rui-Qin MAN
;
Kai KANG
;
Xiao-Yan TANG
;
Hao-Min CHEN
;
Guo-Hua LIU
;
Bian-Li XU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Viral; analysis; immunology; Bunyaviridae Infections; diagnosis; immunology; Cercopithecus aethiops; Female; Fluorescent Antibody Technique, Indirect; Humans; Immunoglobulin G; analysis; immunology; Male; Middle Aged; Orthobunyavirus; immunology; isolation & purification; Sensitivity and Specificity; Vero Cells
- From: Chinese Journal of Preventive Medicine 2012;46(2):165-168
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop an indirect immunofluorescence assay (IFA) for detection of IgG antibodies against new bunyavirus.
METHODSThe 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.
RESULTSThe 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).
CONCLUSIONThe 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.
