1.Molecular inversion probe-based single base mutations of hepatitis B virus resistance gene analysis
Kai CHANG ; Shuangrong JIA ; Feng PAN ; Fake LI ; Feng WANG ; Weiping LU ; Shaoli DENG ; Ming CHEN
Chinese Journal of Laboratory Medicine 2014;(5):337-341
Objective To establish a molecular inversion probe ( MIP) method for detection of single base drug-resistance mutation in Hepatitis B virus ( HBV) gene.Methods The HBV wild type and YVDD mutant strain were isolated by Daping Hospital of the Third Military Medical University.The MIP was designed and applied to detect the HBV drug-resistance YVDD mutation in one case of wild type and one case of YVDD mutant HBV strain isolated previously.The results of MIP method were compared with that of sequencing to evaluate the detection accuracy.Results Thermal cycling single-base extension and connection reaction performed by Taq DNA Ligase and Ampligase DNA Ligase could ensure the specificity of the detection.The optimum probe concentration of MIP was 1 nmol/L.Through detection of the target gene with different DNA concentrations , the detection sensitivity of MIP was determined as 1 nmol/L.The results of MIP were consistent with that of sequencing method in detection of the clinical samples.Conclusion MIP is successfully used to detect single-base drug-resistance mutation in HBV gene.
2.Detection and susceptibility test of suspected 4 414 cases of urogenital tract mycoplasma infection in Chongqing
Shuangrong JIA ; Meng HU ; Linhua JIANG ; Jinmei ZHU ; Shiqiao ZHAO ; Zhongyu CHEN
Chongqing Medicine 2017;46(5):609-611
Objective To investigate the positive rates and susceptibility of Ureaplasma urealyticum(Uu) and Mycoplasma hominis(Mh) in urogenital mycoplasma infection under three years.Methods Culture,identification and susceptibility test were performed on 4 414 specimens collected from suspected patients with mycoplasma infection by using Antu mycoplasma kits.Results In the 4 414 patients,2 295 cases with mycoplasma infection were detected and the positive rate was 51.99%.The infection rates of Uu and Mh respectively were 40.69% and 2.08%,and the both infection rate was 9.22%.Antibiotic sensitive rates of josamycin(JOX),doxycycline(DOX),clarithromycin(CLA),gatifloxacin(GAT) and erythromycin(ERY) were 96.03%,95.51 %,78.69 %%,77.21 % and 72.55 %.Drug resistant rates of roxithromycin(RXT),thiamphenicol (THI),clindamycin (CLI) and clarithromycin(CLA) were 16.90%,22.27%,41.96% and 17.60%.Conclusion Uu is the predominant mycoplasma in urogenital tract infection in the study.DOX,JOS,GAT and ERY can be chosen as the fist line drugs for the treatment of urogenital tract infection.RXT,THI,CLI and CLA with high drug resistant rates are not recommended to be used.