Genetic variation characteristics of HA gene of influenza A virus (H3N2) in Guizhou province from 2017 to 2019 / 中国基层医药

Objective:To analyze the genetic variation characteristics of the HA gene of influenza A virus (H3N2) in Guizhou province from 2017 to 2019. Methods:Twenty strains of influenza A virus (H3N2) were randomly selected from 10 network laboratories in Guizhou province for RNA extraction. Reverse transcriptase-polymerase chain reaction and sequencing were performed. The products were analyzed using bioinformatics software.Results:The nucleotide homology of the HA gene of the 20 strains was 97.7%-100%, which was highly homologous to the vaccine strains A/Hong-Kong/4801/2014 recommended by WHO in 2017 and A/Singapore-INFIMH/16-0019/2016 recommended by WHO in 2018, but they were significantly different from the vaccine strain A/Kansas/14/2017 recommended by WHO in 2019. Genetic analysis showed that the 20 strains were divided into two branches, and the strains that were prevalent in 2019 were located in different branches, with marked genetic differences. Key site analysis showed mutations in antigenic determinants A, B, C, and E and mutations in the anterior and posterior walls of receptor binding sites. Key site analysis also showed that there was an increase in the number of glycosylation sites compared with the vaccine strains prevalent in the same year. Genetic distance, antigen sites, and glycosylation sites were slightly different between virus strains prevalent in 2017-2018 and virus strains prevalent in 2019. Conclusion:The HA gene of the influenza A virus subtype H3N2 in Guizhou province from 2017 to 2019 showed heterogeneity and gene mutation, especially in 2019. Therefore, close monitoring of the genetic evolution of the influenza A virus subtype H3N2 is necessary.

Expression level of serum HBV RNA in HBeAg-positive chronic hepatitis B patients at different periods and its value of measurement / 临床肝胆病杂志

Objective To investigate the expression level and potential clinical value of serum HBV RNA in HBeAg-positive chronic hepatitis B (CHB) patients at different periods. Methods A total of 61 CHB patients who attended the outpatient and inpatient services of Department of Hepatology, Hangzhou Xixi Hospital, from August 2019 to December 2020 were enrolled, and according to the antiviral therapy for HBeAg-positive CHB patients, they can be divided into group A with untreated HBeAg-positive CHB (HBeAg+ and HBV DNA+) patients, group B with treatment-experienced patients before HBeAg seroconversion (HBeAg+ and HBV DNA-), and group C with treatment-experienced patients after HBeAg seroconversion (HBeAg- and HBV DNA-). Peripheral blood HBV RNA load was measured at different periods, and its correlation with HBsAg and HBV DNA was analyzed. The t -test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between groups; a Pearson or Spearman correlation analysis was used to describe the correlation between two variables. Results The positive rates of HBV RNA in these three groups were 100% (22/22), 88.2% (15/17), and 22.7% (6/22), respectively. In group A, HBV RNA was positively correlated with HBsAg and HBV DNA ( r =0.612 and 0.922, both P < 0.01), while in groups B and C, there was no correlation between HBV RNA and HBsAg. Group B had significantly higher levels of HBV RNA and HBsAg than group C ( Z =-4.44 and -2.41, both P < 0.05). The HBV DNA-positive group had a significantly higher level of HBV RNA than the HBV DNA-negative group ( Z =-6.16, P < 0.01). Conclusion After HBV DNA clearance achieved by antiviral therapy with nucleos(t)ide analogues in CHB patients, serum HBV RNA can still be detected in some of these patients. Since HBV RNA only comes from cccDNA in the liver, it can better reflect viral replication activity in the liver than HBV DNA and thus has a certain clinical value in the management of CHB patients.

Proposal for detection of 2019-nCoV nucleic acid in clinical laboratories / 中华检验医学杂志

In December, the outbreak of a novel coronavirus (2019-nCoV) in Wuhan, China, has attracted extensive global attention. On January 20, 2020,the Chinese health authorities upgraded the coronavirus to a Class B infectious disease in the Law of the People's Republic of China on the Prevention and Treatment of Infectious Diseases, and considered it as Class A infectious diseases in disease control and prevention. On January 22, 2020, the 2019-nCoV nucleic acid detection test was listed as the diagnostic criteria in the "guidelines for diagnosis and treatment of pneumonia due to 2019-nCoV (Trial Version 2)" . Therefore, standardizing the operation process of the 2019-nCoV nucleic acid detection in clinical laboratories has become a top priority. It is of paramount importance to establish standard protocols for detection of the 2019-nCoV nucleic acids in clinical laboratories to improve the reliability of the results and ensure the biosafety of laboratory personnel.

Correlation between CD4 Count and HIV-1 Viral Load among ART Naive Patients Attending ICTC, SMS Medical College, Jaipur

Objectives: To study the correlation between CD4 count &HIV-1 viral load among ART Naive patients attending ICTCSMS Medical College, Jaipur.Material and Methods: This study was conducted on 250 HIVserologically confirmed, ART Naive cases from ICTC, SMSJaipur. RNA extraction was done from plasma samples byQiagen Viral RNA Mini Kit then HIV-1 Viral load wasdetermined by Qiagen HIV-1 viral load kit on ABI 7500 Fast dxReal Time PCR, while CD4 count was done on FACSCALIBUR flowcytometer (BD Biosciences). SPSS ver. 21.0was used to determine correlation between CD4 count & HIV-1viral load.Results: Out of 250, 216 (86.4%) cases were found in whichviral RNA was detected. These samples were correlated withtheir CD4 Count. The mean of viral load was 194746.2791 ±550442.61805 IU/ml while CD4 count was 282.7674 ±217.56456 cells/ul. Females were having Avg. Viral load228506.7273 & CD4 count 337.21 and males were found tohave Avg. Viral load 179791.9866 & CD4 count 258.65Conclusion: This study concluded a negativecorrelation between HIV-1 RNA viral load and CD4 count inHIV-seropositive ART naïve patients of this part of the country.Our study confirmed that HIV-1 RNA viral load levels aresignificantly higher in women than in men, but no suchsignificant gender difference in the CD4 count was found.

The application of serum HBV RNA detection in anti-HBV treatment / 中华检验医学杂志

The persistence of covalently closed circular DNA (cccDNA) in the nucleus of liver cells is a key factor that hinders the cure of chronic hepatitis B. However,it is difficult to eliminate cccDNA with existing anti-HBV therapy. Recent studies have found that serum HBV RNA may be a new indicator reflecting the activity of cccDNA in hepatocytes and evaluating the clinical efficacy of CHB patients . This article reviews recent advances in the properties,detection methods,and clinical significance of HBV RNA, particularly the application of antiviral therapy in CHB patients.

The clinical significance of HCV antibody S/CO values in the diagnosis of active HCV infection in cancer patients / 中华检验医学杂志

Objective To investigate the clinical significance of HCV antibody S /CO values in active HCV infection diagnosis in cancer patients .Methods 390 cancer patients were enrolled from Cancer Hospital Chinese Academy of Medical Sciences between January 2013 and April 2015.All of the cancer patients had pathological diagnosis , including 240 males and 150 females, aged from 25 to 83 years old. HCV antibody and HCV RNA levels were detected using the Abbott i 2000 immunity analyzer and Roche LC480 real-time fluorescent quantitative PCR machine , respectively.The relationship between HCV antibody S/CO value and RNA level was analyzed in the group of HCC and non-HCC patients.Results There were obvious statistical differences in age (P＝0.004), gender (P＜0.001) and HCV antibody levels (P＜0.001) between the group of HCC and non-HCC patients.There was no statistical difference in distribution of RNA positive rate between the two groups (P＝0.528).Using ROC curve analysis, the best cut-off value to diagnose active HCV infection in cancer patients is 10.0 with sensitivity 97.6%and specificity 81.3%. According to the results of the ROC curve , the cut-off was 11.4 and 10.4 in HCC and non-HCC patients respectively.Conclusion The best cut-off value to diagnose active HCV infection in cancer patients is 10.0, either in HCC or in non-HCC.

Clinical value of serum HBV RNA / 临床肝胆病杂志

Although there are various indicators for evaluating the effect of anti-HBV therapy,they have low accuracy and sensitivity.New indicators are still needed to guide clinical practice.Recent studies have found that HBV RNA might be a new potential indicator for clinical detection.This article reviews the basic concepts of HBV RNA,related detection methods,and the value of HBV RNA in clinical diagnosis.

Correlation between HCV genotype and anti-HCV antibody level in hepatitis C patients / 中华检验医学杂志

Objective To investigate the relationship between anti-HCV antibody level and hepatitis C virus genotype in the patients.Methods Total of 603 anti-HCV positive serum samples were collected during 2013 to 2014 by retrospective research method.HCV RNA were detected in anti-HCV positive samples by repeat test and the genotype were detected in HCV RNA positive samples.The distribution of anti-HCV level in different hepatitis C genotype patients was analyzed and the body's response to viral antibodies and viral genotype correlation with anti-HCV concentration interquartile range was explored.Rates among genotype groups were compared using chi-square test.Results Totally 412 of 603 (68.33％) samples were anti-HCV positive by double reagent screening.174(42.3％) samples were detected as HCV RNA positive.The distributions of different anti-HCV level in different genotype patients were 1a(n =8) 1/8,1/8,4/8,2/8;1b(n =112)25.9％ (29/112),17.0％ (19/112),25.9％ (29/112),31.3％ (35/112);2a(n =14)3/14,4/14,5/14,2/14;3a(n =11)3/11,6/11,2/11,0/11;3b(n =16)4/16,11/16,1/16,0/16;6a(n =8)2/8,2/8,1/8,3/8 with anti-HCV concentration interquartile range respectively.The anti-HCV concentration distribution was different in patients with different HCV genotypes.The anti-HCV concentration distribution in patients of 1 b,2a and 6a genotypes were evently,while anti-HCV level was relatively high in 1a (13.65) and relatively low in 3b (8.77).There were differences in different genotypes of antibody concentrations (x2 =35.2,P ＜ 0.05).Conclusions There was correlation between anti-HCV level and HCV genotype.Because there were fewer cases in some genotypes,it was necessary to investigate more samples to corfirm the above results.

Improved assembly procedure of viral RNA genomes amplified with Phi29 polymerase from new generation sequencing data

BACKGROUND: New sequencing technologies have opened the way to the discovery and the characterization of pathogenic viruses in clinical samples. However, the use of these new methods can require an amplification of viral RNA prior to the sequencing. Among all the available methods, the procedure based on the use of Phi29 polymerase produces a huge amount of amplified DNA. However, its major disadvantage is to generate a large number of chimeric sequences which can affect the assembly step. The pre-process method proposed in this study strongly limits the negative impact of chimeric reads in order to obtain the full-length of viral genomes. FINDINGS: Three different assembly softwares (ABySS, Ray and SPAdes) were tested for their ability to correctly assemble the full-length of viral genomes. Although in all cases, our pre-processed method improved genome assembly, only its combination with the use of SPAdes allowed us to obtain the full-length of the viral genomes tested in one contig. CONCLUSIONS: The proposed pipeline is able to overcome drawbacks due to the generation of chimeric reads during the amplification of viral RNA which considerably improves the assembling of full-length viral genomes.

Factors Associated with a Low CD4 Count among HIV-1 Infected Patients at Enrolment into HAART in Jos, Nigeria.

Aim: To determine the factors associated with a low CD4 count among HIV-1 positive patients. Study Design: Cross-sectional study. Place and Duration of Study: Adult HIV clinic at the Jos University Teaching Hospital, Jos, between October 2010 and April 2011. Methodology: Data on demographic, clinical and laboratory variables for 218 HIV-1 infected patients aged 20 years and older were analysed. A low CD4 cell count was defined as CD4 cell count <200 cells/ml based on the WHO criteria for severe immune suppression. A multivariate logistic regression modeling was fitted to determine the variables that were independently associated with a low CD4 count. Results: Of the 218 HIV-1 infected patients, 119 (54.6%) had a low CD4 count at enrolment. The odds of having a low CD4 count was: 7 times higher in patients with WHO clinical stage 3 or 4 compared to those with stage 1 or 2 (P<.001) and 4 times higher in those with HIV RNA viral load ≥4.6 log10 copies/ml compared to those with less (P<.001); but the odds of having a low CD4 count was reduced by 63% in those patients that were resident in Plateau State compared to those resident outside the state (P=.01). Conclusion: Our study patients were more likely to have a CD4 count <200 cells/ml which would suggest late presentation/ late HIV diagnosis and thus a delayed opportunity for timely access to HIV care and initiation of antiretroviral therapy. There is the need to intensify efforts in early routine HIV counseling and testing not only in health facilities in the cities but also in smaller towns and rural communities, so as to reduce the frequency of late HIV diagnosis with its potential implications.

Clinical symptoms and courses among men who have sex with men with acute human immunodeficiency virus infection / 中华传染病杂志

Objective To describe the clinical characteristics,CD4+ and CD8+ T cell counts as well as human immunodeficiency virus (HIV) RNA of acute HIV infection in men who have sex with men,and their correlations with the disease progression.Methods One hundred cases of acute HIV infection were followed up.Nuclear acid sequence-based amplification (NASBA) was used for plasma HIV RNA screening.Flow cytometry was served to test the CD4+ and CD8+ T cell counts.Hepatitis B surface antigen (HBsAg) and anti-hepatits C virus (HCV) antibody were detected using enzyme-linked immunosorbent assay.Rapid plasma reagin was applied to screen for Treponema pallidum antibody.Antibody positive specimens were tested with Treponema pallidum particle assay for validation.The positive results were identified as infection.Results Ninety-six cases were aged between 20 and 50 years old.Among 100 cases,9 were HBsAg-positive; 4 were anti-HCV positive; 40 were co-infected with syphilis.During the follow-up period,the median CD4+ T cell counts in the 1st and 3rd month were 510/μL and 499/μL,respectively.The median HIV RNA in 1st,3rd,6th and 24th month were 4.37,4.00,4.31 and 4.43 lg copy/mL,respectively.CD8+ T cell counts did not show significant change during the study.Among 38 rapid progressors,the initial mean CD4+ T cell counts was (358.0± 134.6)/μL,which was significantly lower than that of the non-rapid progressors with a mean CD4+ T cell counts of (559.2±203.4)/μL.Meanwhile,the initial mean HIV RNA of the rapid progressors was 4.71 lg copy/ mL,while that of the non rapid progressor was 4.18 lg copy/mL.The initial CD8+ T cell counts of the rapid progressors and non rapid progressors were 1 250.1/μL and 1 247.2/μL,respectively.Conclusions Acute HIV-1 infected men who have sex with men tend to be young.The initial CD4+ T cell counts and HIV RNA during acute infection could be used to predict the disease progress.

Detection of the specific antigens and viral RNA in the tissues of the BALB/c mice infected with hantavirus / 中国医师杂志

Objective To establish an evaluation system about animals infected with hantavirus,an observation of the BALB/c mice infected with hantavirus was made.Methods BALB/c mice were infected with hantavirus by intramuscular injection with stock solution.The specific antigen from BALB/c mice tissues after 3 days was detected with enzyme-linked immunosorbent assay (ELISA) and viral RNA with real-time polymerase chain reaction (RT-PCR).Results Within a short term,the specific antigen and viral RNA were detected from the brain and liver at day 3 after infection,but not be detected from the heart,spleen,lung,and kidney samples.Conclusions The results provided ones with some information on animals infected with hantavirus.

Analysis of the results of external quality assessment for hepatitis C virus RNA tests / 中华检验医学杂志

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.

Development of a novel quantitative real-time assay using self-reporting duplex mutation primers for detection of HCV / 中华检验医学杂志

Objective To establish a novel real-time PCR method to detect HCV RNA using Selfreporting duplex mutation primers.Methods The recombinant vector pMD18-T-HCV 5′-NCR was used as the calibrator.The Self-reporting duplex mutation primers were designed according to the gene sequence.And then the PCR reaction system was optimized and evaluated.The specificity,sensitivity and reproducibility of real-time PCR were estimated,The serum specimens from 90 cases(30 cases of HCV,30 cases of other viral hepatitis and 30 healthy volunteers) were tested with this real-time PCR; Results were compared with those obtained using a commercial TaqMan kit.Results The assay was established.It showed linearity over a wide range from 20 - 109 IU/ml.Intra-experimental coefficients of variation(CVs) were 1.37% -4.59%,and inter-experimental CVs were 1.58% -4.81%,respectively.There was no significant difference of HCV genome number tested by the two methods(R2 = 0.95) in 30 hepatitis C patients; HCV DNA was not detected in any serum samples of 30 healthy volunteers by the two methods.The specificity was 100%(60/60).All the samples in patients with clinically confirmed HCV infections showed HCV RNA positive.There wass good correlation between the quantitaive results and results obtained using the commercial TaqMan kit.Conclusions It is demonstrated that real-time PCR is a reliable,accurate and feasible assay for HCV.The establishment of this assay provided alternative technology for clinical diagnosis or therapeutic drug monitoring in the field of HCV infection and epidemiologic survey.

The correlation between APOBEC3G mRNA in peripheral blood mononuclear cells and serum hepatitis C viral RNA level / 中华传染病杂志

Objective To study the relationship between APOBEC3G mRNA level in peripheral blood mononuclear cells (PBMC) and serum hepatitis C viral RNA level in patients with chronic hepatitis C infection. Methods TaqMan real-time fluorescence relative quantitative polymerase chain reaction (RT-PCR) was used to quantify APOBEC3G mRNA levels in PBMC from 49 patients with chronic hepatitis C (CHC) and 31 healthy subjects. The relationship between APOBEC3G mRNA level and hepatitis C virus (HCV) viral load was analyzed. SPSS11. 0 statistics software was used for t test and regression analysis. Results APOBEC3G mRNA level in CHC patients [(1.5×10-5±1.9×10-5 ) copy/mL] was significantly lower than that [( 5. 2 × 10-5 ± 5. 5 × 10-5 ) copy/mL] in the healthy control subjects (t=-3.005, P＜0.01). While APOBEC3G mRNA level was not related with HCV viral loads (r=-0.082, P＞0.05). Conclusion HCV has an inhibitive effect on APOBEC3G expression, whereas APOBEC3G doesn't affect HCV replication directly in vivo.

External quality assessment for the quantitative detection of HCV RNA in China / 中华检验医学杂志

Objective Anti-RNase virus-like particles containing HCV RNA 5'-UTR were used as positive samples in national external quality assessment ( EQA ) to evaluate the competency of clinical Laboratories for the quantification of HCV RNA and analyze the possible problems of domestic kits. Methods The quality control samples with target values in EQA panels were distributed nationally twice by National Center of Clinical Laboratory (NCCL) to participating laboratories for the quantification of HCV RNA in 2008 and 2009. Each panel consisted of 5 samples. All participants were required to carry out the detection and to return results in expected time. Positive samples were virus-like particles which had been calibrated against the WHO HCV International Standard (NIBSC96/798)and the results of positive samples from participants should be in the range of target value of logarithm ± 0. 5. The 2nd panel in 2008 contained the common HCV genotypes and the 2nd panel in 2009 contained serial diluted samples of genotype 1b. The results of positive samples detected with 3 different lots reagent (21001,21078 and 21097) from the 2nd EQA in 2009 were statistically analyzed using the analysis of variance, then Dunnett'S T3 and Tamhane'S T2 were used if heterogeneity of variance was found. Results There was 390 participating laboratories in 2008 and 428/426 in 2009. The percentages of laboratories within the range of target value of logarithm ± 0. 5 for varied genotypes were different. The percentages of laboratories for 1b were more than 91%, for 2a were 93.7% and 74. 2% ,for6 were 83.3% and 80. 3%. The CVfor the low-level sample was higher than that for the high-level sample in the same year. The numbers of laboratories reporting false-negative samples in 2008and the 2nd in 2009 were 5, 1 and 10 respectively. Statistical differences were found among the results of four quality control serum samples using 3 different reagents( F = 288.23, 324. 79, 291.98 and 261.16,P ＜0. 01 ). Conclusion The competency for detecting low concentration samples and samples with genotype 2a or 6 needs to be improved.

Comparison and quality control between domestic and foreign hepatitis virus nucleic acid amplification technology reagents for quantitative and qualitative tests / 中华检验医学杂志

Hepatitis virus NAT reagents are now widely used clinically. However, the qulity of domestic and foreign NAT reagents varies dramatically. The main reasons for these differences including the manufacture technique, test principle and assay procedure were discussed in this paper and current status of the quality control of the NAT reagents were also described. Finally, it was pointed out that strengthening public supervision and laboratory internal control are very important for the quality improvement of the domestic reagents.

The tracability and uniform unit of hepatitis C virus RNA quantification by domestic made real-time fluorescence quantitative PCR method / 中华检验医学杂志

Objective To determine the conversion equation for X(copies/ml, lg) quantity values of domestic HCV RNA quantitative fluorescence amplification assays approved by SFDA and Y( IU/ml, lg)reference values of standard substance. Methods The second generation WHO International Standard (NIBSC code:96/798) was mixed with human AB blood type serum to create 7 different dilutions which included 100 000, 50 000, 25 000, 10 000, 5 000, 2 500 and 1 000 IU/ml. Two different batches of each three domestic hepatitis C virus RNA real-time fluorescence quantitative PCR assays and 2 different batches of each assay were employed to detect the 7 different concentration samples with real-time PCR. Each test was performed 4 times repeatedly. Results The correlations between X( copies/ml,lg) values of domestic HCV RNA assays and Y(IU/ml,lg) reference values of standard substance were as follow,Assay A:Y =0. 902 0 X+0.284 9,R2 =0.953 3,P＜0. 01,n =56;Assay B: Y=0. 875 7 X +0.562 4,R2 =0.956 5,P＜0.01,n =56; Assay C: Y = 0. 843 8 X + 0. 560 5, R2 = 0. 945 8, P ＜ 0. 01, n = 56. Conclusions All the conversion equations are different among the quantity value of three assays and the reference values of standard substance, that suggests it is necessary to perform more stringent traceability analysis for the quantity values of 3 assays. Through standardizing the quantity values preliminarily, the conversion equation can enhance the comparability between the quantity values of different assays, and provide a standard of HCV RNA virus load detection for clinical diagnosis and treatment monitoring of HCV infection.

Study on the correlation between anti-thyroid autoantibodies and hepatitis C virus infection / 中华传染病杂志

Objective To explore the correlation between anti-thyroid autoantibodies and hepatitis C virus (HCV) infection. Methods Four hundred and sixty-two samples with positive thyroid peroxidase antibody (TPOAb) and (or) thyroglobulin antibody (TgAb) were collected. Three hundred and eighty age and gender matched subjects with negative TPOAb and TgAb were selected as controls. The anti-HCV antibody was examined in all the cases using the third-generation enzyme-linked immunosorbent assay (ELISA), HCV RNA qualitative examination was examined further in those who had positive anti-HCV antibody. Meanwhile, 195 subjects with hepatitis C, 150 healthy subjects and 150 subjects with hepatitis B were tested for thyroid-related markers. The data were analyzed by independent-sample t test and chi square test. Results The HCV infection rate in 462 thyroid autoantibodies positive subjects was 1.30% and 0.53% in 380 thyroid autoantibodies negative subjects. There was no significant difference of the HCV infection rate between two groups (X2=1.322, P>0.05). In the subjects with hepatitis C, 30.8% were TPOAb positive, 30.8% were TgAb positive, which were significantly different from those of healthy subjects and subjects with hepatitis B (X2=21.496,X2=30.454;P<0.01). Conclusions HCV infection rate does not increase in subjects with abnormal thyroid autoimmunity. However, positive rate of thyroid autoantibodies increases in subjects with hepatitis C, which suggests that thyroid-related markers should be examined in hepatitis C patients.

Detection of hepatitis E virus RNA by real-time fluorescent RT-PCR / 中华检验医学杂志

Objective To investigate the clinical significance of detection of hepatitis E virus (HEV) RNA in sera from patients with acute hepatitis E using real-time reverse transcription (RT)-PCR to detect hepatitis E virus RNA in sera from patients with acute hepatitis E.Methods A real-time RT-PCR assay, which can amplifies and detect the conserved region on ORF3, was used in this study. 434 outpatients and hospitalized patients with acute HEV infection was enrolled into this study.Simultaneously,the serum samples from 40 patients with HAV infection, 100 patients with HBV infection and 110 healthy blood donors were collected as the control The real-time RT-PCR was performed to detect HEV RNA in all these sera.Results 232 sera (53.5%) were positive for HEV RNA by real-time RT-PCR and all of the control were negative.The results of real-time RT-PCR and anti-HEV IgM (ELISA) were concordant in 67.1% samples.There was significant difference between the two methods ( Kappa = 0.308, P = 0.000 ).The first serum sample from five serum samples of the patients was positive for HEV RNA and negative for anti-HEV IgM.Follow-up studies showed all the five sera samples were positive for anti-HEV IgM.HEV RNA in serum could be detected between 2 and 10 days.Conclusions The real-time fluorescent RT-PCR method has high specificity, and can be applied to the qualitative detection of the serum with genotypes Ⅰ and Ⅳ of hepatitis E virus.Its clinical use can improve the early diagnosis of HEV.