Comparative evaluation of susceptibility testing methods for colistin and polymyxin B on isolates of Enterobacteriaceae / 中华检验医学杂志

Objective:To provide a promising and optimal laboratory susceptibility-testing method for the clinical usage of antibiotic (polymyxin), four susceptibility-testing methods were performed and the broth microdilution (BMD) was chosen as the gold standard.Methods:A total number of eighty-eight nonduplicate clinical Enterobacteriaceae specimes were collected from January to December of 2019 in the First Affiliated Hospital, Fujian Medical University. Among the clinical specimens, of which six strains were positive for mcr-1. The minimal inhibitory concentration (MIC) of polymyxin of the clinical specimens were examined by the following methods: (1) broth microdilution, (2) colistin broth disk elution, (3) Vitek-2?, (4)BD PhoenixTM,(5)commercial broth microdilution. With BMD as reference, essential agreement (EA), categorical agreement(CA), very major error(VME) and major error (ME) of polymyxins for different methods were analyzed. The Kappa-consistency testing, paired Chi-square testing and the Spearman-rank correlation testing were used to analyze the consistency between the four antimicrobial susceptibility testing methods and the gold standard.Results:Taking broth microdilution as reference, the EA of colistin broth disk elution, Vitek-2?, BD PhoenixTM, commercial broth microdilution were 94.32% (83/88), 92.05% (81/88), 90.90% (80/88), and 96.59%(85/88), respectively. The CA of all the four methods were 100% (88/88). No VME and ME were recorded for four methods. Moreover, the consistency between four susceptibility testing methods and the gold standard is acceptable (Kappa values=1, P<0.001, McNemar test P=1 and r>0.5, P<0.05). Conclusions:In the present work, four susceptibility testing methods all met the standards recommended jointly by the Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing, of which the performance of the commercial broth microdilution and CBDE fared relatively well. Thus, these four methods could be routinely used in clinical microbiology laboratory of our hospital for colistin and polymyxin B susceptibility testing.

The genetic polymorphism of CYP7A1 in Fujian Han HBV infected patients / 中华检验医学杂志

Objective The study aims to investigate the associationbetweencholesterol 7α-hydroxylase (CYP7A1) gene polymorphism and different clinical outcomes after Hepatitis B virus (HBV)infection in Fujian Han population and lay a foundation for understanding the mechanisms of genesis anddevelopment of HBV-related diseases.Methods Case-control study was conducted.586 patients of HBVpersistent infection without antiviral therapy and 225 HBV rehabilitation patients (35-55 years old) werecollected from May 2015 to June 2016 in the Liverish Center of First Clinical College of Fujian MedicalUniversity.The group of HBV persistent infection without antiviral therapy included 246 patients with chronichepatitis B, 177 patients with hepatitis B-related cirrhosis, and 163 patients with hepatitis B-related liver cancer.The rs3824260, rs4738687and rs8192871 loci of CYP7A1 gene were detected by improved multipleligase detection reaction (iMLDR).Logistic regression analysis and chi-square test were used to analyze thegenotyping results.Results Three SNPs ( single nucleotide polymorphisms ) of CYP7A1 gene wereselected and compared between HBV persistent infection group and HBV rehabilitation group and betweenchronic hepatitis B subgroup, liver cirrhosis subgroup and liver cancer subgroup.After adjustment for factorsincluding age andgender, there was no significant difference in the distribution of rs3824260 genotype amongthe groups(χ2 =1.565,P =0.459), however,the frequency of allele C in HBV rehabilitation group wassignificantly higher than in HBV persistent in fectiongroup for men (χ2 =4.365,P =0.037), whereas thefrequency of rs3824260 CC and CT was more likely to be observed in liver cancer group than in non -livercancer group (chronic hepatitis B subgroup and liver cirrhosis subgroup ) for women (χ2 =5.768,P =0.012;χ2 =10.130,P =0.001).The frequency of rs4738687 GG genotype was more likely to be observed innon-liver cancer group than in liver cancer group (χ2 =4.403,P =0.041;χ2 =6.940,P =0.009).Theresults of gender stratification showed that there were significant differences in the distribution of rs 4738687among the HBV persistent infection groups for men (χ2 =10.697,P =0.030), however, there was nosignificant difference in the distribution of rs4738687 among the HBV persistent infection groups for women(χ2 =4.627,P =0.329), and there was no significant difference in the distribution of genotype frequencyand allele frequency among all groups(χ2 =0.489,P =0.792).There was no significant difference after sexstratification either (χ2 =1.282, P =0.526;χ2 =1.565,P =0.465) .Conclusions These findingssuggested that CYP7A1 gene polymorphism was related todifferent clinical outcomes in Fujian Hanpopulation.The rs3824260 mutation had a certain gender preference and the mutation allele was detected ina higher proportion in male patients.Male HBV patients with rs3824260 C allele had more chance ofswitching to rehabilitation.The rs4738687 was likely to be related to the occurrence of liver cancer in FujianHan population, and GG genotype may delay the occurrence and development of liver cancer especially in themale group.The rs8192871 was not found to be related to the different clinical outcomes of HBV infection.

Epidemiological and molecular virological characteristics of HBV infected patients of hospital with simultaneously positive HBsAg and anti-HBs / 临床检验杂志

Objective To investigate the epidemiological and molecular virological characteristics in HBV-infected patients with copositive HBsAg and anti-HBs.Methods HBV serological markers were analyzed in 52 070 specimens.The epidemiological characteristics of HBsAg and anti-HBs simultaneously positive patients (the experimental group) and HBsAg positive and auti-HBs negative patients (the control group) were compared.The S protein of HBV coding region was amplified by semi-nested PCR and sequenced.The statistical differences between the two groups were compared in different gene regions,genotypes and different clinical diagnosis.Results HBsAg was positive in 20.40％ (10 621/52 070) of all specimens.In the patients with positive HBsAg,2.48％ (263/10 621) was positive anti-HBs.The prevalence of co-positive HBsAg and auti-HBs was higher in aged 0 to 9 years and greater than or equal to 80 years than that in other age,and the prevalence of positive HBsAg and negative anti-HBs was completely opposite.The mutation rate of S protein in the experimental group was significantly higher than that in the control group (1.52％ vs 0.81％,P ＜0.01) with the mutation in the major hydrophilic region (MHR) (1.68％ vs 0.57％,P ＜0.01).The mutation rates of S protein of HBV carriers,chronic hepatitis B (CHB) patients and patients with liver cirrhosis (LC) in the experimental group were significantly higher than those in the control group (1.47％ vs 0.65％,1.28％ vs 0.84％,2.21％ vs 0.44％,P ＜0.05,respectively),except for the patients with hepatocellular carcinoma (HCC) (1.97％ vs 2.21％,P ＞ 0.05).Conclusion Co-positive HBsAg and anti-HBs in HBV-infected patients was more common in HBsAg positive patients aged 0 to 9 years and greater than or equal to 80 years than the others.Coexistence of HBsAg and anti-HBs in HBV-infected patients may relate to immune escape caused by mutation of S protein (mainly MHR).The mutation rates of S protein in the two groups of patients,co-positive HBsAg and anti-HBs and the positive HBsAg combined with negative anti-HBs,were associated with the stage of liver disease.

The relationship between HBV P region genetic muation and serum markers in hepatitis B patients in Fuzhou region / 中华检验医学杂志

Objective To investigate the correlation of hepatitis B patients in Fuzhou between resistance patterns in HBV P region and genotype,HBeAg,the hepatitis B process.Methods This was a retrospective study.The serum and clinic data of 1 115 hepatitis B patients were collected from the inpatient and outpatient Center for Liver Diseases in First Affiliated Hospital of Fujian Medical University between October 2011 and January 2015.HBV DNA was extracted and sequenced using the Sanger method to detect HBV genotype and resistance mutations in P region,HBeAg and HBeAb concentration were detected by chemiluminescent assay.The relationship between P region resistance mutations pattern,HBV genotype,serum HBeAg and the hepatitis B process was analyzed.The x2-test was used to compare the resistance rate and positive rate.Results There were significant differences between 14 kinds of resistance loci and the genotype distribution(x2 =30.788,P =0.004),the C/B genotype ratio of three common resistance loci (rtM204V/I,rtL180M,rtA181T/V) were 85/82,49/25 and 27/9,respectively,which in genotype C was higher than genotype B.The resistance ratio of hepatocellular carcinoma,liver cirrhosis,chronic hepatitis B,hepatitis B carriers was 31.4％ (11/35),37.6％ (65/173),27.3％ (146/535) and 21.8％ (43/197),respectively,which showed significant difference between the four clinical diagnosis (x2 =11.858,P =0.008).The highest percentage of resistance was liver cirrhosis,followed by hepatocellular carcinoma and chronic hepatitis B.There was significant difference in the distribution of HBV genotype between HBeAg (+) group and HBeAg (-) group (x2 =11.093,P =0.001),the HBeAg positive rate in genotype C [37.53％ (295/786)] was higher than in genotype B[35.62％ (280/786)].However,the total resistance rate between HBeAg (+) group and HBeAg(-) group was not significantly different[23.7％ (136/573) and 24.6％ (52/211),respectively,x2 =0.07,P =0.791].Conclusions HBV genotype was related to the resistance rates,HBeAg levels and the progress of hepatitis B.The resistance rate and HBeAg positive rate of genotype C were higher than those of genotype B,and clinical outcomes were worse in genotype C.HBV resistance rates and HBeAg levels were related to the progress of hepatitis B,the higher the resistance rates,the worse clinical outcomes.

Studies on the association of single nucleotide polymorphisms of HLA-DP and DQ genes with the outcome of chronic hepatitis B virus infection / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of single nucleotide polymorphisms in the HLA-DP and DQ genes with the outcome of chronic hepatitis B virus infection.</p><p><b>METHODS</b>Two hundred and four healthy subjects, 255 clearance subjects, 204 asymptomatic HBV carriers (AsC), 136 chronic hepatitis B (CHB), 68 liver cirrhosis (LC) and hepatocellular carcinoma (HCC) were enrolled. Genotypes of rs3077, rs9277535 and rs2647050 were determined by sequence specific primers-PCR (PCR-SSP).</p><p><b>RESULTS</b>By using healthy subjects and clearance subjects as the control groups, rs3077 and rs9277535 were significantly associated with chronic HBV infection under additive and dominant models (P< 0.05). Meanwhile, haplotypes GGA, AGA, AAA appeared to be protective factors against chronic HBV infection (P < 0.05). By using AsC as the control group, comparison with the CHB, LC and HCC groups showed no association of the 3 SNPs or haplotypes with the clinical outcome (P > 0.05).</p><p><b>CONCLUSION</b>HLA-DP gene polymorphisms are strongly associated with chronic HBV infection. The presence of A allele at rs3077 and rs9277535 of the HLA-DP gene may decreased the risk for chronic HBV infection.</p>

An innovative HBV genotyping and drug-resistant mutation detection platform / 中华检验医学杂志

Despite decades of the prevention and control.China is still one of the countries with high prevalence of hepatitis B virus (HBV).It is imperative for us to innovate.Lab diagnostic platforms for drug-resistant mutation and HBV genotyping to enhance the management level in patients with CHB.Specifically,the following three strategies should be included:firstly,choose an appropriate platform for drug-resistant mutation detection according to the actual situation in laboratory.Secondly,establish a practical and simple genotyping platform for real-time monitoring of HBV genotypes.Thirdly,combined with clinical,explore the correlation among genotypic resistance,phenotypic resistance and clinical resistance when different medications are adopted,confirm the percentage or threshold of resistant strains causing clinical resistance,and ascertain the influence of HBV genome polymorphism (genotype) on clinical prognosis.These strategies above mentioned will lay a foundation for CHB personalized treatment and prognosis.

Advances in HBV drug-resistance mutations and detection technologies / 中华检验医学杂志

Drug-resistance mutation is inevitable during long-term monotherapy with nucleus(t)ide analogues in hepatitis B patients.Early detection the drug-resistance mutations could contribute to adjusting the therapeutic regimen,reducing resistance rate,improving therapeutic efficacy and performing individual treatment.