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Resistance mutation during anti-HBV and anti-HCV treatment:question and strategy

Chinese Journal of Laboratory Medicine.2013;(3):193-200. doi:10.3760/cma.j.issn.1009-9158.2013.03.001

Both long term follow up and real world research on chronic hepatitis B have accumulated data on nucleos (t) ide analogues in anti-HBV treatment,which shows that resistance mutation turns out to be the major obstacle in achieving response.Identification of genotypic resistance at early stage is key to improve strategy.Deep-sequencing will be helpful to predict resistance earlier.Direct acting autiviral agents on hepatitic C virus tell us resistance at the very beginning,however,more data is still needed to elucidate how to use resistance monitoring of anti-HCV treatment.

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The immunological and clinical significance of IgM in primary biliary cirrhosis

Shanshan HUANG ; Xiong MA

Chinese Journal of Laboratory Medicine.2013;(3):201-205. doi:10.3760/cma.j.issn.1009-9158.2013.03.002

Primary biliary cirrhosis (PBC) is a slowly progressive autoimmune disease of the liver.Studies on the pathphysiology of this disease have demonstrated the immune-mediated destruction of the intrahepatic bile ducts and portal infammation seem to be the result of an intense autoimmune response directed against the autoantigens located in the biliary epithelium.Immunologically,PBC is characterized by the presense of antimitochodrial antibodies,elevated serum IgM and lymphoid infiltration in the portal tract.Indeed,the relationships between IgM and PBC have been a major focus of research on its pathphysiology,which lead to an increased understanding of the abnormal immune responses involved in PBC.This review summarizes data on the biological and clinical significance of Hyper-IgM in PBC,and highlights the potential role of IgM for the diagnosis and treatment of PBC.

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Research progress in immunopathogenesis and detection of hepatitis B

Yu CHEN

Chinese Journal of Laboratory Medicine.2013;(3):208-211. doi:10.3760/cma.j.issn.1009-9158.2013.03.004

HBV infection is the key factor that causes liver inflammation,liver injury and immune dysfunction.Chronic hepatitis caused by HBV,a non-cytotoxic virus,is a kind of immune injury induced by liver cells that clear virus infection in immune system.Effector T lymphocyte plays a central role in immune system to clear HBV in liver cells,and the outcome of HBV infection is directly influenced by its functions and quantity.After antiviral therapy for patients with sustained HBV infection,whether to restore and rebuild immune function is critical in achieving ideal end point of treatment.Technologies,mainly including stimulation and culture of HBV antigen,flow cytometry,enzyme-linked immunoassay to detect cytokines,etc,are used in detection of HBV-specific T lymphocytes.

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Cancer biomarkers and their clinical application for hepatocellular carcinoma diagnosis

Meng FANG ; Chunfang GAO

Chinese Journal of Laboratory Medicine.2013;(3):212-216. doi:10.3760/cma.j.issn.1009-9158.2013.03.005

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in China because of high incidence of hepatitis B virus (HBV) infection.HCC is diagnosed at a late stage in most of the cases; therefore the prognosis of patients with HCC is generally poor.Early diagnosis and surgical treatment are of great clinical desirable to improve prognosis of HCC.Tumor marker is an effective means for early diagnosis,prognosis assessment and recurrence monitoring.In addition to alpha fetoprotein (AFP),Lens culinaris agglutinin-reactive AFP (AFP-L3),des-γ-carboxy prothrombin (DCP),glypican-3,N-glycome markers,candidate-susceptibility genes,microRNAs and several other biomarkers have been revealed as potential HCC markers and will be applied in clinical laboratory gradually.In this review,the efficacies of novel HCC markers and their possible implications for clinical application are described.

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Analysis of suspicious results of serum HBV DNA detected by fluorescence quantitative PCR

Zhanguo CHEN ; Wu ZHOU ; Zhongyong WANG ; Yalei JIN ; Zhihua TAO

Chinese Journal of Laboratory Medicine.2013;(3):217-221. doi:10.3760/cma.j.issn.1009-9158.2013.03.006

Objective To analyze the suspicious results of serum HBV DNA by fluorescence quantitative PCR and develop appropriate countermeasures in order to improve the quality of detection of HBV DNA.Methods Blood samples of patients from the First Affiliated Hospital of Wenzhou Medical College from 2008 to 2011 were analyzed for HBV DNA by fluorescence quantitative PCR.1969 cases of suspicious results,judged by the rule of review the results of serum HBV DNA combined with the historical results,PCR amplification curve,HBV serum markers and clinical diagnosis,were analyzed and redetected by using of two different reagents,careHBV PCR Kit and careHBV PCR Kit V2,at the same time.The consistency and inconsistency ratio of the results were evaluated.Both the reasons of inconsistent and the undetected rates of careHBV PCR Kit were analyzed.The two reasons for the inconsistent results included the reagent related factors,e.g,showing no amplification curve caused by the false negative and abnormal low efficiency of amplification curve,and the non reagent related factors such as operating pollution and other sample factors.Results There were 115 154 blood samples were detected for HBV from 2008 to 2011 and 1969 samples (1.71％) with suspicious results were redetected.The consistency and inconsistency results were 1588 (80.65％) and 381 (19.35％),respectively.Every year from 2008 to 2011,the percentage of the inconsistent results caused by the reagent related factors were 18.87％,20.23％,51.33％ and 59.57％ respectively,which showed an increasing trend,and the percentage of inconsistent results caused by the nonreagent related factors were 81.13％,79.77％,48.67％ and 40.43％ respectively,which showed a declining trend year by year.The undetected rates of careHBV PCR Kit were 2.49％,4.08％,10.09％ and 14.47％ respectively,showing an increasing trend.Conclusions The redetection for the specimens with the suspicious results by using of different reagents can avoid the blind detection of HBV DNA and reduce the experimental error.All the clinical samples for quantitative HBV DNA including the mutations of HBV gene can be measured accurately and effectively,which is helpful to hepatitis B patients for antiviral therapy.

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Analysis of the results of external quality assessment for hepatitis C virus RNA tests

Rui ZHANG ; Lunan WANG ; Kuo ZHANG ; Jiehong XIE ; Yu SUN ; Jinming LI

Chinese Journal of Laboratory Medicine.2013;(3):271-276. doi:10.3760/cma.j.issn.1009-9158.2013.03.020

Objective To evaluate the performance of HCV RNA detection in the first EQA program in 2012 and analyze possible problems in clinical laboratories.Methods The panel consisting of 5 samples was distributed to 927 laboratories.Each panel contains one negative sample and 4 positive samples,which were virus-like particles calibrated by international standard.The pere ent agreements of all the laboratories for qualitative and quantitative results were calculated.Genomic means (GM) and standard deviations (s) of all laboratories and each reagent were calculated.The overall GM and the GM of each reagent were compared with expected results and correlation curves were calculated.Results The percent agreements of sample 1211,1212,1213,1214 for qualitative results wcrc 99.5％ (403/405),98.5％ (400/406),100.0％ (405/405),100.0％ (406/406),respectively.The percent agreement of the negative sample was 99％ (401/405).The percent agreements of sample 1211,1212 and 1213 for quantitive results were similar,which were 93.8％ (549/585),92.3 ％ (541/586) and 94.5％ (554/586).However,the agreement of sample 1214 was only 87.7％ (514/586)and the agreement of sample 1214 for reagent A was 67.2％ (92/137).The overall GM agreed with expected results,while GMs of reagent C,E and G deviated from expected results.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent C were 4.22,3.56,5.16 and 5.90,respectively.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent E were 4.52,3.78,5.55 and 6.29,respectively.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent G were 4.83,4.36,5.72 and 6.56,respectively.Conclusions The overall results of HCV RNA qualitative and quantitative detection are satisfactory.However,some problems still exist,such as deviation of GM of some reagents,the interassay variability,systematic deviation and accidental deviation,which show that the quality of reagents should be improved.

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Mechanisms of carbapenem resistance in Enterobacteriaceae and the strategies for control of carbapenem-resistant Enterobacteriaceae

Ding LIU ; Liping ZHANG

Chinese Journal of Laboratory Medicine.2013;(4):300-302. doi:10.3760/cma.j.issn.1009-9158.2013.04.003

Monitoring data of antibiotic susceptibility from both home and abroad showed that carbapenem-resistant Enterobacteriaceae (CRE) are being on the rise,which represents an increasingly urgent clinical danger and an emerging public health concern worldwide.In order to effectively prevent CRE transmission in healthcare settings,we should not only pay high attention to the capability building of the laboratory of clinical microbiology,but also actively explore into the mechanisms of carbapenem resistance in Enterobacteriaceae.What's more,we should substantially raise the detection capacity of laboratory on CRE,effectively carry out the antimicrobial stewardship program,and strictly implement hand hygiene and contact precautions.(Chin J Lab Med,2013,36:300-302)

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Multilocus sequence typing of carbapenem resistant Acinetobacter baumannii from Zhejiang Province

Qing YANG ; Zhihu ZHOU ; Yiqi FU ; Hua ZHOU ; Yan CHEN ; Yunsong YU

Chinese Journal of Laboratory Medicine.2013;(4):303-307. doi:10.3760/cma.j.issn.1009-9158.2013.04.004

Objective To investigate the molecular epidemiology of carbapenem resistant Acinetobacter baumannii from Zhejiang Province using multilocus sequence typing.Methods Three hundred and two imipenem or meropenem resistant A.baumannii isolates were collected from eleven tertiary hospitals of eleven different regions of Zhejiang Province in 2009 and 2010.Multilocus sequence typing was used for molecular typing.eBURST was used to analyze the results of multilocus sequence typing.PCR was used to detect the OXA-type carbapenemase genes.Results Seventeen sequence types (STs) were identified from three hundred and two A.baumannii isolates.eBURST analysis revealed eleven STs belonged to clonal complex 92 (CC92) and corresponded to European clone Ⅱ lineage.CC92 was the predominant carbapenem resistant A.baumannii clone of Zhejiang Province,which accounted for 94.4％ (285/302) of all isolates and distributed in all 11 hospitals.blaOXA-23 gene was identified in 97.4％ (294/302) of all isolates.Conclusions Carbapenem resistant A.baumannii CC92 were clonal disseminated among multiple hospitals of Zhejiang Province.blaOXA-23 was the most popular carbapenemase gene in carbapenem resistant A.baumannii.(Chin J Lab Med,2013,36:303-307)

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Meta-analysis of pyrosequencing for the rapid detection of isoniazid-resistance in Mycobacterium tuberculosis

Qi GUO ; Jue LI ; Zhongyi HU ; Ruijuan ZHENG

Chinese Journal of Laboratory Medicine.2013;(4):329-332. doi:10.3760/cma.j.issn.1009-9158.2013.04.009

Objective To evaluate the accuracy of pyrosequencing for the mutation detection of katG gene in isoniazid resistance in Mycobacterium tuberculosis using Meta-analysis.Methods Searching PubMed,Web of Science,Elsevier,and China National Knowledge Infrastructure (CNKI),Weipu and WANFANG DATA to obtain the relevant English and Chinese-language articles,respectively.A written protocol and explicit study selection criteria was followed.Quality of included trials was assessed by QUADAS (quality assessment of diagnostic accuracy studies).Subsequently,the characteristics of the included articles were appraised and extracted.Heterogeneity of the included articles was tested by using STATA 10.0,which was used to select proper effect model.The fixed effects model was adopted using Meta-Disc software.Finally,sensitivity analysis was performed.Results Totally 114 research papers were collected and 9 articles were selected.The accordance between pyrosequencing and conventional sequencing result was 100％.Eight studies were involved including 945 specimens when katG gene was detected.The overall sensitivity and specificity were 0.77 (0.73,0.80) and 1.00 (0.99,1.00),respectively.The area under the SROC was 0.9882.As inhA gene was detected,the overall sensitivity and specificity were 0.19 (0.15,0.24) and 1.00 (0.98,1.00).The test was stable.Conclusions Our meta-analysis reveals that pyrosequencing is a highly specific tool for detection mutation of katG gene of isoniazid resistance.This result suggests that it is useful for screening of isoniazid resistance in diagnostic test.(Chin J Lab Med,2013,36:329-332)

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How to strengthen communication between department of microbiology and the clinical departments

Yanping LUO ; Jiyong YANG

Chinese Journal of Laboratory Medicine.2013;(4):375-376. doi:10.3760/cma.j.issn.1009-9158.2013.04.020

With the rapid development of clinical laboratory technology and clinical improvement,the discordance between department of microbiology and clinical departments got more and more.Based on the analysis of interface issues existed between them,we discussed and explored how to do a wonderful job to communicate with each other.(Chin J Lab Med,2013,36:375-376)

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