Study on the efficacy evaluation of real-time fluorescence assay of RNA isothermal amplification in the treatment of patients with positive pulmonary tuberculosis / 天津医药

@#Objective To investigate the value of real-time fluorescence detection technique of RNA (SAT) constantamplification in monitoring the effect of chemotherapy on patients with sputum positive pulmonary tuberculosis. Methods Sixty-two patients were selected, who were clinically diagnosed as the first-time retreatment for sputum smear-positive pulmonary tuberculosis and were hospitalized in our department from June 2015 to December 2016. After two-month standard anti-tuberculosis treatment, sputum samples were detected by Peng’s vessel acid-fast staining, BACTEC MGIT- 960 culture and strain identification, SAT detection. The BACTEC MGIT-960 culture and strain identification were used as gold standards, the value of SAT during the monitoring the therapeutic effect of the anti-tuberculosis drugs was assessed. Results After the treatment, 49 cases out of 62 showed positive results in mycobacterium tuberculosis culture test, among them 42 patients were diagnosed as human type mycobacterium tuberculosis, 7 patients were diagnosed as nontuberculous mycobacteria infection, and 13 cases showed negative results in mycobacterium tuberculosis culture. Thirty-two cases showed positive results in sputum Peng’s vessel acid-fast staining test, and 30 cases showed negative results. Forty-one cases showed positive results in SAT test and 21 cases showed negative results in SAT. SAT results were well concordant with sputum culture results (Kappa value=0.964), and the sensitivity, the specificity, the positive predictive value and the negative predictive value of SAT were 97.62%, 100%, 100% and 95.24% respectively. Peng’s vessel acid-fast staining results were badly concordant with MGIT-960 culture results (Kappa value=0.086), and the sensitivity, the specificity, the positive predictive value and the negative predictive value of Peng’s vessel acid-fast staining were 54.76%, 55.00%, 71.88% and 36.67% respectively. Conclusion SAT results can be used as good indices during the monitoring therapeutic effects of drugs used for the first-time retreatment in patients with sputum smear-positive pulmonary tuberculosis, which is worth promoting.

Diagnostic value of detecting Mycobacterium tuberculosis rRNA via SAT in anti-TB treatment / 实用医学杂志

Objective To study the clinical value of simultaneous amplification and testing for detection of Mycobacteria tuberculosis(SAT-TB)in sputum samples and bronchoalveolar lavage fluid(BALF)samples. Methods Totally 169 sputum samples and 151 BALF samples from suspected pulmonary tuberculosis patients were detected by both SAT and Bactec MGIT960.The sensitivity,specificity,positive predictive value(PPV)and negative predictive value(NPV)of the samples using SAT-TB were calculated. Results Taken the results of BD960 as the reference,the sensitivity,specificity,PPV and NPV using SAT-TB of sputum samples were 84.00% (42/50),93.06%(67/72),89.36%(42/47)and 89.33%(67/75)respectively;and those of BALF samples 89.19% (33/37),95.12%(39/41),94.29%(33/35)and 42.39%(39/92)respectively.Taken clinical diagnostic results as reference standard,the sensitivity,specificity,PPV and NPV using SAT-TB of the sputum samples were 57.73% (56/97),93.06%(67/72),91.80%(56/61),and 62.04%(67/108)respectively;and those of BALF samples 51.82%(57/110),94.29%(39/41),96.61%(57/59)and、42.39%(39/92)respectively.The sensitivity,specificity, PPV and NPV using BD960 of the sputum samples were 51.55%(50/97),95.83%(69/72),94.34%(50/53),and 59.48%(69/116)respectively;and those of BALF samples 33.64%(37/110),90.24%(37/41),90.24%(37/41) and 33.64%(37/110)respectively.Conclusion SAT-TB is a rapid and sensitive method for the detection of Myco-bacteria tuberculosis in sputum and BALF samples.It can improve the detection rate of mycobaterium tuberculosis.

Isothermal nucleic acid amplification technology and its application in molecular identification of Chinese materia medica / 中草药

Accurate identification of varieties is the most important part of quality control of Chinese materia medica (CMM). To find an efficient, convenient, and accurate identification method is the development trend of identification technology of CMM. Compared with the traditional identification method based on phenotypic markers, DNA molecular marker technology is more accurate and reliable, suitable for the identification of closely related species and sample with confusion and multiple sources, but unable to realize the rapid identification due to the limits of the PCR technology, such as high cost, complex procedures, and the drawback of long time. Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification technology, with the advantages of high specificity, high sensitivity, simple, rapid, low cost, etc., become another new technology after PCR to realize the rapid molecular identification of CMM successfully. In this paper, the common isothermal amplification of nucleic acid technology and its application in the study of molecular identification of Chinese herbal medicines were reviewed analysis, to provide a reference for the study of rapid molecular identification system of Chinese herbal medicines.

Nucleic acid detection technology used in blood screening of blood donors / 国际检验医学杂志

Objective To evaluate the necessity and feasibility of nucleic acid test for donors blood screening .Methods From July 1 ,2011 to December 31 ,2014 ,a total of 170 316 blood samples which were negative in enzyme-linked immunosorbent assay (ELISA)and qualified in aianine aminotransferase detection ,were selected in this study stochastically .All the samples were detected hepatitis B virus(HBV) ,hepatitis C virus(HCV) ,human immunodeficiency virus(HIV) by nucleic acid amplification technology (NAT) .NAT positive samples were reconfirmed in National Center for Clinical Laboratories(NCCL) .Results A total of 160 cases of nucleic acid reactive samples were found out ,the total response rate was 0 .09% ,The response rate of Roche nucleic acid detec-tion system was 0 .10% ,response rate of David nucleic acid detection system was 0 .08% ,there was no significant difference be-tween the two methods(P>0 .05) .In 27 cases of specimens ,14 cases were confirmed as HBV DNA positive ,no HCV RNA and HIV RNA were detected ,the confirmed positive rate was 51 .85% .There were 2 samples detected by chemiluminescence HBsAg reactivity .Conclusion ELISA screening of blood donors has missing phenomenon ,nucleic acid detection method could be used as an effective supplement of the ELISA ,could improve the safety of blood for clinical use ,detection sensitivity is better than ELISA .

Analysis of HCV positive plasma and manufacturing of HCV RNA national standard candidate / 대한수혈학회지

BACKGROUNDS: Standardization of nucleic acid amplification techniques (NAT) which can be achieved by the use of standard to validate reproducibility and sensitivity in each assay run is necessary before the introduction of such methods for routine screening of blood and blood products for viral contaminants. The objective of this study was to analyze the serological and genotypic characteristics of HCV positive plasmas and to manufacture the HCV RNA national standard candidate. METHODS: We obtained three plasmas from Blood Transfusion Research Institute, Korea, with highly positive HCV RNA plasmas (#37, #40, #46) and with normal plasma for dilution. All the plasmas were confirmed by enzyme immunoassay (EIA) test for anti-HIV, HBsAg, anti-HCV and by polymerase chain reaction(PCR) for HBV DNA, HIV RNA, HCV RNA. The genotypes of those were confirmed by INNO-LiPA HCV II. HCV RNA national standard candidate was manufactured by dispensing the diluted plasma into about 2,000 vials. Each vial was rapidly frozen using liquid nitrogen and was kept in refrigerator at -70 degrees C. RESULTS: All plasmas were identified as anti-HIV, HBsAg, HBV DNA, and HIV RNA negative plasmas. The genotypes of those were confirmed as 1b for #37, 1b or 2 for #40 and 2a or 2c for #46, respectively. Sample #37 was selected as the candidate material. After manufacturing, we obtained 1,944 vials for the candidate. CONCLUSION: In this study, we analyzed HCV positive plasmas and manufactured the HCV RNA national standard candidate. In near future, this material would be established for national standard to increase in the safety of blood and blood products in Korea.