摘要
目的通过荧光定量PCR(fluorescent quantitative PCR,FQ-PCR)技术对血浆病毒灭活前后乙型肝炎病毒DNA(HBV-DNA)浓度进行测定,判断亚甲蓝(MB)光化学法灭活乙型肝炎病毒的效果,为临床判断病毒灭活效果提供直接和客观依据。方法用4份已证实为HBV-DNA阳性的血浆,经可见光(>3000LUX)照射不同时间(0、5、10、15、30min)后,用FQ-PCR分别测定这些血浆的HBV-DNA浓度,并与未用MB处理的血浆作比较,判断随照射时间不同,HBV被灭活的效果。结果4份HBV-DNA阳性患者的血浆未经处理时,其病毒核酸浓度分别为5.6×107、3.2×106、1.8×106和3.5×105拷贝/ml,加入MB未经照射时HBV-DNA浓度即明显下降(P<0.05),可见光照射5min后HBV-DNA浓度分别下降为未经任何处理时的浓度的15.89%、11.56%、4.39%和1.23%。随照射时间的延长,病毒核酸浓度逐渐下降,照射30min后,HBV-DNA浓度均下降为零。结论用MB加光照30min处理,可达到将血浆乙肝病毒灭活的效果。
Objective To evaluate the effect of methylene blue and visible light by FQ-PCR assay on the inactivation of plasma hepatitis B virus,and to establish a direct and impersonal basis for the surveillance of cural inactivation. Methods Four samples of plasma of HBV-DNA positive were added with methylene blue. Then the plasma was processed with visible light radiation（30 000 Lux） for 0, 5, 10, 15,30min respectively. Moreover,the concentration of HBV-DNA samples was quantitatively detected by the method of FQ-PCR and compared with untreated samples to evaluate the effect of HBV inactivation and its relationship with the time of radiation. Results The original concentration of HBV DNA was 5.6×10^07 , 3.2 ×10^6 , 1.8×10^6 ,3.5×10^5 copies/ml respectively. After being added with methyene blue, the concentration of HBV DNA was significantly decreased（P〈0.05）. And after 5 rain of radiation, the concentration decreased to 15.89%,11.56%,4.39% and 1.23% in the proportion of the original concentration. After 30 rain of radiation, the concentration was up to zero. Conclusion The plasma HBV could be inactivated by methylene blue and visible light radiation for 30 min.
出处
《国际检验医学杂志》
CAS
2006年第1期1-2,6,共3页
International Journal of Laboratory Medicine
关键词
肝炎病毒
乙型
DNA
病毒
聚合酶链反应
消毒
Hepatitis B virus
DNA,virus
Polymerase chain reaction
Disinfection
