| Objective To elaborate HCV genotype distribution of the main monitoring population(DUS, MSM, FSW, STD, DRV, MPO, PRG, YST) in HIV sentinel surveillance in Anhui Province using cross-sectional study. In the meanwhile, we analysis and comparison above different groups with HCV genotype distributions whether there are differences. In addition, The HCV genotype differences were also explained in socio-demographic characteristics such as gender, age, and infection route among different groups in Anhui Province, providing a scientific basis for the analysis of the spread and prevalence of HCV.Methods In this study, cross-sectional study was adopted. We selected Wuhu, Anqing, Suzhou, Fuyang, Hefei five prefecture-level city as for investigation points from south, central and north region. According to the National HIV sentinel surveillance program, each sentinel adopt the snowball(DUS and MSM) stratified proportional sampling and dynamic monitoring(FSW) dynamic monitoring(STD, DRV,PRG) network recruiting(among MSM) and multi-stage stratified cluster sampling(YST) these five different sampling methods for monitoring population screening from April 1 to June 30 in 2012-2014. In this study, the number of 6,098 were screened in Wuhu and anti-HCV positive was 37 from which 28 serum samples were collected; The number of 4,221 were screened in Anqing and anti-HCV positive was 24 from which 22 serum samples were collected; The number of 2,216 were screened in Suzhou and anti-HCV positive was 18 from which 13 serum samples were collected; The number of 3,784 were screened in Suzhou and anti-HCV positive was 48, from which 35 serum samples were collected; In Hefei 9,700 respondents were screened and anti-HCV positive was 291, from which 265 serum samples were collected. All of the 363 HCV antibody positive serum samples from 2012-2014 were incorporated into this study, which they would be detected by RT-PCR, then the amplified products were sequenced and used for gene alignment and typing. All the monitored populations recruited were investigated by anonymous questionnaire after the implementation of informed consent. Besides, 16 anti-HCV positive plasma samples were collected in hospital outpatient and inpatient from Chuzhou and Hefei, and registered their basic information.The method of serological tests include HCVRNA extraction and amplification; Molecular biology methods include nucleic acid amplification products were sequenced and HCV genotyping. Determination of sequence assembly using Contig Express / Chromas Pro and Bio Edit software; Gene sequencing assembling through the neighbor joining of Mega6.0 software then carried out gene sequence alignment with the standard strain from Genbank,to build phylogenetic tree of HCV NS5 B and CE2 region.Epi Data 3.1 and SPSS17.0 were used to establish a database and manage the data. Frequency distribution, normality test, chi-square test test and One-Way ANOVA were used to analyze the data.Results 1. A total of 26,219 cases was screened and 418 cases of HCV antibody-positive from the main monitoring population in HIV sentinel surveillance, including DUS 1770 people(anti-HCV positive 286), FSW 4648 people(anti-HCV positive 39), STD 4866 people(anti-HCV positive 32), MSM 2871(anti-HCV positive 21), DRV 813 people(anti-HCV positive 2), MPO 3209 people(anti-HCV positive 23 people), PRG 4000 people(12 positive anti-HCV), YST 4042 people(anti-HCV positive 3), antibody positive rate was 16.16%(DUS), 0.84%(FSW),0.66%(STD), 0.73%(MSM), 0.25(DRV), 0.72%(MPO), 0.30%(PRG) and 0.07%(YST). In this study, 362 serum samples and 16 plasma samples were collected. With all population of HCV antibody positive samples for nucleic acid detection, nucleic acids were detected 214 positive(DUS 152, FSW 22, STD 17, MSM 9, MPO 9, PRG4 and DRV1), YST undetectable HCV nucleic acid positive. Nucleic acid detection rate was 57.36%(DUS), 70.59%(FSW), 76.19%(STD), 66.67%(MSM), 100%(DRV), 45.00%(MPO), 44.44%(PRG) and 0.00%(YST). Sixteen anti-HCV positive plasma samples were detected eight Nucleic acids positive.2. The results show, there are eight separate HCV genotypes and three kinds of mixed genotypes in DUS population, and the proportion of each type were: 3b subtype 40.13%, 3a subtype 21.71%, 1b subtype 11.84%, 6n subtype 9.21%, 6a subtype 6.58%, 6u subtype 3.29%, 2a subtype 2.63%and 1a subype 0.66%, mixed genotypes 1b / 3b, 2a / 3b and 3a / 3b each account for 1.32%; There are five HCV genotypes in FSW population, namely the proportion of genotype 1b subtype 56.52%, 6a subtype 21.74%, 3a subtype 13.04%, 2a and 3b subtype, were account for 4.35%; There are three HCV genotypes in MSM population, and the proportion of each type were:1b subtype 75.00%, 2a and 6a subtype has the same proportions, account for 12.50%; STD population also were detected three HCV genotypes, the proportion of each subtype were: 1b subtype 70.59%, 6a subtype 23.53% and 6n subtype 5.88%; DRV population were detected only 1 HCV RNA positive samples, that was genotype 1b; MPO population were detected two HCV genotypes, the proportion were 1b type 55.56% and 2a 44.44%, respectively; PRG population were detected four cases, 1a subtype 2 cases, accounting for 50.00 %, 3a and 6a each were detected in one case, account for 25.00%; GP population also were detected two kinds of HCV genotypes, 1b subtype were 75.00% and 2a subtype 25.00%. The study did not find genotypes 4, 5 and 7.3. One hundred and forty-nine samples from DUS population which correctly genotyped with 100 males cases mainly popular genotype was 3b subtype, followed by 3a type, accounting for 38.00% and 22.00%. And genotype 6 and 1 also accounted for a large proportion were 21.00% and 15.00 respectively. Besides, mixed genotypes were detected only one sample. Forty-nine female samples also mainly subtype 3b, followed by 3a subtype, and the proportion of these two genotypes accounted for 44.90% and 22.45%. Besides, subtype 6 had a certain proportion. Three male and five female patients with hepatitis C main genotype was 1b, followed by a 2a type. By the Fisher’s exact probability test, HCV genotype groups was no significant difference in this two population between male and female in both the gender distribution(F= 9.589, P >0.05).4. Five genotypes from DUS population on age distribution as follows: <45-year-old age group of the proportion of different HCV genotypes was 3b(37.21%), 3a(22.48%), 6 type(20.16%) type 1(13.18%) and mixed(4.65%), respectively; And 2a subtype had a smallest proportion(2.33%).The proportion of HCV genotype in more than 45 years old group were, 3b type(60.00%), 3a type(20.00%), type 6(10.00%) and type 1(10.00%), mixed genotypes in this age group is not detected. HCV genotype differences in the overall distribution of the two age groups was statistically significant(χ2 =6.159, P>0.05); But patients who infected genotype 3 have a higher proportion in both age groups, and the mixed genotypes found only in lower age groups. In less than 45-years-old age group,FSW, STD, MSM population main genotype was 1b(28 samples), followed by 6 type(10 samples), 2a, 3a and 3b subtypes were detected 2, 3 and 1 samples, respectively. HCV main genotype in more than 45 years age group was only 1 type(3 samples) and 6 type(1 sample), type 2 and 3 was not found in this age group. In less than 45-years-old age group, DRV, MPO, PRG and GP population HCV genotype distribution mainly was genotype 1b(7 samples), followed by 2a subtype(3 samples), genotyes 3a and 6 were not found. HCV main genotype in more than 45years age group was only 1b(7 samples) and 2a(3 samples) subtypes, type 3 and 6 in this age group were not found. It was can be seen that genotype 1 have a higher proportion in two age groups of seven categories population except DUS population. Only genotype 6 had a higher proportion in FSW, STD and MSM population of the two age groups; But genotypes 2 and 3 in the high age group was not detected out; Genotype 2a have a higher proportion in two age groups of DRV, MPO, PRG and GP population, and the genotype 3 and 6 in the high age group were not found. Age(measurement data) meet normal distribution, using the single-factor analysis of variance, the difference was not statistically significant(F = 1.012, P> 0.05).5. Two hundred and nineteen cases of anti-HCV positive population, 68.04% have a history of drug abuse spread, 21.92% for sexual transmission, 10.05% from other routes of infection such as blood transfusions and other unexplained. It showed that there were significant differences HCV genotype distribution in the different routes of infection(F = 98.046, P <0.01). Transmission route of drug use main genotype were 3(3a / 3b) and 6(6a / 6n / 6u), accounting for62.42% and 18.79%, respectively, followed by subtype 1(1a / 1b)(12.75%), and subtype 2(2a) occupies a smaller proportion(2.01%). Transmission through sexual contact, including FSW, MSM and STD population, they mainly genotype was1-type(69.57%), followed by 6-type(22.92%), type 2 and type 3 had a certain proportion, accounted for 4.17% and 8.33%, respectively. Other routes of infection include blood transfusion and unexplained, mainly genotype were type 1(63.64%) and 2(27.27%), and type 3(4.55%) and 6(4.55%) each account for a smaller proportion.6. After electrophoresis, sequencing and phylogenetic analysis, the results showed: The HCV NS5 B zone electrophoresis showed a positive number was 216; By HCV C / E2 zone electrophoresis showed a positive number was 178. The two districts were detected 172 similarly, 165 samples of two regions detection had a consistent genotyping results that can determine the genotype; The overall classification identical rate was 95.93%(165/172); In addition, seven samples measured genotyping results were inconsistent which mixed genotypes were 1b / 3b(2 samples), 3a / 3b(3 samples) and 2a / 3b(2 samples).Conclusions The main monitoring population in HIV sentinel surveillance in Anhui Province had eight kinds of HCVsingle subtypes(1a, 1b, 2a, 3a, 3b, 6a, 6n and 6u). Anhui Province has not yet to found genotype 1a, 3 and 6 in related literature by now. In addition, this study did not find genotypes 4, 5 and 7.Different populations had significantly different genotype: DUS population predominant HCV genotype was 3b; Sexual contact group(FSW, MSM, STD) predominant HCV genotype were 1b and 6(6a, 6u and 6n); General population(MPO and GP) was majority of genotype 1b and 2a.HCV genotypes had no significant gender differences, but the differences of their age distribution were more obvious.Different infection routes had significant difference genotypes. |
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