[Comparison of direct immune-fluorescent assay and real-time quantitative PCR in detecting the Hantavirus].
OBJECTIVE
To compare the differences between the direct immuno-fluorescent assay (DFA) and real-time quantitative PCR in detecting the Hantavirus (HV) in rat lungs.


METHODS
From April to October in 2012, a total of 479 rats were caught by mouse-trap in residential or wild areas in Huxian, Jingyang, and Meixian of Shaanxi province, where haemorrhagic fever with renal syndrome (HFRS) was highly prevalent. The rats were dissected to take the two lungs, one was frozen and applied immuno-fluorescent assay to detect HV antigen while the other one was extracted its RNA and detected HV nucleic acid by real-time quantitative PCR. Then we compared the positive rate of the two methods.


RESULTS
Out of the 479 rats, 105 were caught from residential areas and the other 374 were caught from wild areas. Among the 105 rats caught from residential areas, no HV were detected out neither by DFA nor by real-time quantitative PCR. Among the 374 wild rats, 13.1% (49/374) were detected HV positive by DFA and 14.7% (55/374) were detected HV positive by real-time quantitative PCR. The difference showed no statistical significance (χ(2) = 0.402, P = 0.526). When detecting each lung sample, the HV positive rate was 10.2% (49/479) under the detection by DFA while the HV positive rate was 11.5% (55/479) under the detection by real-time quantitative PCR. The difference had no statistical significance (χ(2) = 1.286, P = 0.257) and the consistency coefficient was 68.2% under the paired chi-square test analysis, which showed high consistency (u = 11.759, P < 0.05). The sensitivity of real-time quantitative PCR to detect HV was 77.6% (38/49) comparing with DFA as standard, and the specificity was 96.1% (413/430). Out of the 9 suspected HV positive sample detected by DFA, 6 were confirmed positive by real-time quantitative PCR and 3 were denied.


CONCLUSIONS
Compared with the DFA, real-time quantitative PCR could also be used to detect the infection of HV in rats, and the result might be more stable.
Laboratory for Molecular Virology, Shaanxi Center for Disease Control and Prevention, Xi'an 710054, China.
Peng-bo
PB
Yu
Shen
S
Li
Jing
J
Wei
Chang-an
CA
Ma
Xiao-ling
XL
Lu
Shui-quan
SQ
DU
Lu-yuan
LY
Guan
Yuan
Y
Zheng
Jian-hua
JH
Dong
Chao-feng
CF
Ma
Jing-jun
JJ
Wang
D000818:Animals; D019085:Fluorescent Antibody Technique, Direct; D006476:Orthohantavirus / Q000302:isolation & purification*; D006480:Hemorrhagic Fever with Renal Syndrome / Q000453:epidemiology / Q000517:prevention & control; D008168:Lung / Q000821:virology; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction