[Establishment and application of fluorescent quantitative PCR method for detecting human herpes virus type 6].
OBJECTIVE
To establish fluorescent quantitative PCR method for detecting human herpes virus type 6 (HHV6).


METHODS
According to the specific sequence of human herpes virus type 6 genes, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The fragment generated from HHV 6 gene as template was cloned into the pMD18-T vector which was constructed from the pUC 18. And the positive recombinant plasmid was 1:10 diluted and used as standard quantitative template to make the standard curve for sample detection.


RESULTS
The standard curve indicated the linear relationship between Ct (cycle threshold) and template concentration. The clinical samples from 135 cases were detected by this system, 16 cases among 135 were positive.


CONCLUSIONS
The fluorescent quantitative PCR method for the detection of human herpes virus type 6 is simple and accurate, and this method may be helpful to clinical diagnosis.
Dong
Xue-jun
XJ
Central Laboratory, Shaoxing People's Hospital, Shaoxing, China.
Shi
Chang-gen
CG
Qu
Shi-wen
SW
Liu
Zhong-min
ZM
Fu
Jia-ping
JP
D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D015654:Herpesvirus 6, Human / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D016553:Purpura, Thrombocytopenic, Idiopathic / Q000175:diagnosis / Q000821:virology*; D015203:Reproducibility of Results; D019349:Roseolovirus Infections / Q000175:diagnosis / Q000821:virology*; D012680:Sensitivity and Specificity; D055815:Young Adult
D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral