Abstract：In order to improve the detection rate for rabbit HEV RNA, specific primers and TaqMan probes were designed and evaluated in this study. Fourteen complete genome sequences of rabbit HEV isolates downloaded from GenBank were aligned to get the conserved areas in ORF1 and ORF3. Subsequently, TaqMan probes and pairs of primers were designed and synthesized according to the conserved areas respectively. The probe and primers for ORF1 were named as group C and ORF3 as group B, while those had been reported by Jothikumar N were named as group A. Then three groups of different serial 10-fold dilutions of standard plasmid stocks were constructed. The standard curves were generated, and then 49 feces and 44 sera collected from rabbits were tested by the three groups of probes and primers. The results were compared and analyzed. Two newly-designed TaqMan probes and primers for rabbit HEV detection were established and the slopes of the standard curves were -3.455 and -3.469. The positive rates of the three groups (A, B and C) for detection of the 49 rabbit feces were 67.35% (33/49), 67.35% (33/49), 57.14% (28/49) and the mean viral load of HEV RNA (log copies/g) was 6.18, 5.90 and 6.11, respectively. In addition, the positive rates of serum HEV RNA were 65.90% (29/44), 56.82% (25/44) and 50.00% (22/44) and the mean viral load of HEV RNA (log copies/mL) was 3.62, 3.43 and 3.03. Though varied in genomic structure, the application of universal TaqMan probes and primers for rabbit HEV detection does not affect its results.
曾航, 张玉林, 王麟, 刘鹏, 王玲. 实时荧光定量PCR对兔戊型肝炎病毒(HEV)的检测及评估[J]. 中国人兽共患病学报, 2015, 31(7): 612-617.
ZENG Hang, ZHANG Yu-lin, WANG Lin, LIU Peng, WANG Ling. Detection and evaluation of rabbit HEV by quantitative real-time PCR. Chinese Journal of Zoonoses, 2015, 31(7): 612-617.
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2015, Vol. 31 
