Semi-quantitative measurement of HCV RNA by Europium-labeled gene probe
- VernacularTitle:铕标记基因探针半定量检测丙型肝炎病毒RNA
- Author:
Mingzhe NING
;
Mingqing TONG
;
Shiyang PAN
;
Peijun HUANG
;
Junhong WANG
- Publication Type:Journal Article
- Keywords:
Europium;
Chelating agents;
Polymerase chain reaction;
Hepatitis C
- From:
Chinese Journal of Laboratory Medicine
2003;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a semi-quantitative method for measurement of HCV RNA by use of primers and probe, which was sensitive and designed by ourselves, a new europium fluorescent chelate BHHCT. Methods 44 serum samples of HCV infected patients and 20 samples of the healthy people were collected. HCV RNA in serum sample was extracted by HCV fluorescence PCR diagnostic kit produced by Zhongshan University DAAN Gene Co Ltd, and amplified by RT-PCR in which one PCR primer was pre-labeled with biotin. The amplified products were hybridized with capture probes pre-fixed on the microplate. The biotin in the amplified products was conjugated with europium labeled streptavidin. So europium was linked to the target DNA. Then the fluorescent of europium was measured. Results The linear range of this assay was 10 - 10 copies/ml. Both sensitivity and specificity were 100%. Conclusion Europium labeled RT-PCR assay is a sensitive, specific, fast and non-radioactive contaminant method for the measurement of HCV RNA.
