Abstract: Objective To establish a high specificity and sensitivity of detection of infant diarrhea stool specimens in the gut of rotavirus （RV） nested polymerase chain reaction （PCR）,and to explore its preliminary application in clinical diagnosis. Methods according to the RV conservative district gene sequences and USES Primer5.0 software design two pairs of primers of nested PCR to establish a high specificity and sensitivity of detection of RV nested polymerase chain reaction （PCR）,and to evaluate its specific degrees and sensitivity. 136 clinical samples were tested with the established nested PCR method,and the test results were verified by single rt-pcr,colloidal gold and gene sequence analysis. Results the RV specific bands of 192bp were detected by nested PCR,and the other enteroviruses were negative and specific. The RV of 8.705fg/ul can be detected at least by nested PCR method. The detection results of nested PCR and single rt-pcr were 100%,but lowerthan the colloidal gold method,and the difference was statistically significant （χ²=2.615,P<0.05）. The RV products amplified by the nested PCR method were sequenced by nucleotide. By BLAST comparison,the homology of more than 20 known sequences,such as RV and EU679386,DQ870492 and AF531912,was 98%~100%. Conclusion the detection method of nested PCR for intestinal virus （RV） was established,which was characterized by high specificity,high sensitivity and rapid,and worthy of clinical application.

Key words: Intestinal rotavirus, Nested PCR, Set up, Application