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.

Objective To analyze Blimp1 expression difference in the latent and active of tuberculosis patients and the healthy , and assess possibility of as the new tuberculosis diagnostics molecules .Methods 60 patients with active TB (active tuberculosis group) ,50 participants with latent(latent tuberculosis group) ,and 50 healthy people(control group) were enrolled separately .Using fluorescence quantitative polymerase chain reaction were used to determin Blimp 1 mRNA expression in the peripheral blood mono-nuclear cells .Results Blimp1 mRNA expression level of the active tuberculosis group was 15 .35 times than the control group ,and 2 .21 times than the latent tuberculosis group ,the difference was statistically significant (P<0 .05) .Conclusion Blimp1 gene proba-bly plays a role in the immune response to tuberculosis .it provides new ideas for the laboratory diagnosis of tuberculosis and new clues for further exploring the pathogenesis mechanism of tuberculosis .

Objective To establish a new method for rapid detection of β-thalassemia by investigating six clinical common mutation types.Methods Fifty cases of clinical wild-type samples and 42 cases ofβ-thalassemia samples were collected, and β-globin gene was amplified by PCR.Uniform ligation probe ( ULP) specific probes were designed for hybridization reaction to increase the reaction specificity and real-time PCR was performed to increase the sensitivity.After that, PCR products were verified by agarose electrophoresis.After examining the specificity and sensitivity, Kappa test between LDR-ULP method and reverse dot blot( RDB) method was conducted.Results Hybridization efficiency was improved 2.53 times by LDR-ULP hybridization.Each mutant type showed a significant amplification curve, whereas the wild-type had no significant curve within 40 cycles.The limit of determination of this method was 5 pg.The results of 92 cases of peripheral blood samples detected by the method of LDR-ULP and RDB were completely consistent.Conclusion In this study, a simple, inexpensive, rapid new method to detect β-thalassemia were established.

Objective:To reveal the significance of microRNA 155 (miR-155) level or the suppressor of cytokine signaling 6 (SOCS6) level in distinguishing the differentiating tuberculosis (TB) infection.Methods:A case-control study was conducted. A total of 60 patients were enrolled in the study retrospectively, including 20 patients with active pulmonary tuberculosis, 20 patients with latent tuberculosis and 20 patients with other pulmonary infectious diseases (non-TB infection), who visited The Third Affiliated Hospital of Third Military Medical University from January to June of 2017. The expression level of miR-155 and SOCS6 in peripheral blood mononuclear cells (PBMC) of from these patients were examined by using the quantitative real-time polymerase chain reaction (qPCR) methods. The associated statistics and graphs was utilized to obtain the relationship, which were reflected by the Co-index receiver operating characteristic(ROC) curve or calculating the area under ROC curve (AUC), between the miR-155 and SOCS6 in differentiating tuberculosis infection by using the Logistic Regressive analysis methods, MedCalc and GraphPad Prism 8.0 software.Results:Neither of the three index miR-155 (AUC=0.663, P=0.031), SOCS6 (AUC=0.708, P=0.002) and Co-index (AUC=0.718, P=0.001) was outstanding to distinguish the tuberculosis infection and non-TB infection. Moreover, the miR-155 (AUC=0.867, P<0.001) and Co-index (AUC=0.875, P<0.001) were similar sufficient ( Z=0.142, P=0.887) to distinguish the active and latent infections. The Co-index (AUC=0.923, P<0.001) was better ( Z=2.586, P=0.010) than SOCS6 (AUC=0.723, P=0.007), and similar ( Z=1.585, P=0.113) to miR-155 (AUC=0.835, P<0.001) on the distinguishing active and non-TB infection. Conclusions:By performing the qPCR and the correlation-analysis, miR-155 has been considered as a potential biomarker for differentiating latent tuberculosis infection from active tuberculosis infection. Conjoint analysis of miR-155 and SOCS6 benefits the distinguishing active TB infection from other pulmonary infectious diseases.