Erman Daloğlu Aylin, Parkan Ömür Mustafa, Erdoğan Ali, Peker Bilal Olcay, Can Sarınoğlu Rabia, Sağlık İmran, İnan Dilara, Kuloğlu Mehmet Murat, Mutlu Derya, Öngüt Gözde, Çolak Dilek
University of Health Sciences Antalya Training and Research Hospital, Medical Microbiology Laboratory, Antalya, Turkey.
Erciyes University Faculty of Medicine, Department of Medical Microbiology, Kayseri, Turkey.
Mikrobiyol Bul. 2021 Jan;55(1):30-40. doi: 10.5578/mb.20108.
Genotype distribution of hepatitis C virus (HCV) can vary over the years between different patient groups and regions. The prevalence of intravenous drug users (IVDU) is known to increase in our country, yet there are a limited number of studies investigating the distribution of HCV genotypes in this group. These data are essential for monitorization of the changes in HCV epidemiology. The present study aimed to evaluate the five-year results of HCV genotyping among patients infected with HCV related to IVDU and unrelated to drug use. Plasma samples of 720 patients (HCV antibody, HCV RNA positive), which were sent to our laboratory for HCV genotyping between January 2014-March 2019 were analyzed. HCV RNA extraction from plasma samples was performed in the automated-extraction system of EZ1 advanced (Qiagen, Germany) using the EZ1 virus mini kit v2.0 (Qiagen, Germany). Amplicons were obtained by amplifying the 5'NCR and core gene region in the Rotorgene 6000 real-time PCR (Qiagen, Germany) device with the HCV RNA real-time quantitative 2.0 (NLM, Italy) kit. For the genotyping, a commercial line probe assay (LIPA) based on in vitro reverse hybridization GEN-C2.0 kit (NLM, Italy) which can distinguish 1, 2, 3, 4, 6 genotypes and 1a, 1b, 2a/c, 2b, 3a, 3b, 3c, 3k, 4a, 4b, 4c/d, 4e, 4f, 4h, 5a, 6a/b, 6g, 6f/q, 6m, 7a subtypes of HCV, based on variations in the 5'-NCR and core regions was used. HCV genotype distribution of 266 IVDU (93.2%: male; median age: 25 ± 6.82) and 454 non-drug users (51.3%: male; median age: 56.5 ± 16.06) were examined. In order of frequency in the group with IVDU; genotype 1a, 3a, 1b, 4c/d, 2b, 4, 3 were observed and genotype 1, 2a/c and mixed genotype (1+3a) were detected in one patient. In the group without IVDU, in order of frequency; genotype 1b, 1a, 3a, 1, 2a/c, 4 were observed and genotype 2b, 4c/d, 5a, 6a/b, 6 and mixed genotype (3+4) were detected in one patient. Genotypes 1a and 3a were significantly higher in the IVDU group (p< 0.00001, p< 0.00001), while 1b was significantly higher in patients without IVDU (p< 0.00001). Genotypes 1a and 3a were more common in young men (p< 0.00001, p= 0.000163), while 1b was higher in middleaged women (p< 0.00001). The incidence of genotypes 1b (p= 0.021) and 3a (p= 0.012) was higher in foreign nationals than the Turkish patients. When the HCV genotype distribution was examined by years, it was observed that the percentages of genotype 1b and 1a were decreasing, while the percentage of genotype 3a was increasing. As a result, in this study, HCV genotype distribution among IVDU was observed to be different from the general population without IVDU. It was found that genotypes 1a and 3a were more common in the IVDU group. As in the other regions of our country, genotype 1b was found most frequently in the general population. Genotype 3a increases significantly compared to years. In our study, the determination of genotypes existing in different parts of the world may be due to the foreign nationals living in our city and our region is a tourism center. It is also necessary to investigate whether there is an increase in IVDU over the years.
丙型肝炎病毒(HCV)的基因型分布在不同患者群体和地区可能会随时间而变化。已知我国静脉注射吸毒者(IVDU)的患病率有所上升,但针对该群体中HCV基因型分布的研究数量有限。这些数据对于监测HCV流行病学的变化至关重要。本研究旨在评估与IVDU相关和与药物使用无关的HCV感染患者的HCV基因分型五年结果。对2014年1月至2019年3月间送至我们实验室进行HCV基因分型的720例患者(HCV抗体、HCV RNA阳性)的血浆样本进行了分析。使用EZ1病毒小提试剂盒v2.0(德国Qiagen公司)在EZ1 advanced自动提取系统(德国Qiagen公司)中从血浆样本中提取HCV RNA。在Rotorgene 6000实时PCR(德国Qiagen公司)仪器中,使用HCV RNA实时定量2.0试剂盒(意大利NLM公司)扩增5'NCR和核心基因区域以获得扩增子。对于基因分型,使用基于体外反向杂交的商业线性探针分析(LIPA)GEN-C2.0试剂盒(意大利NLM公司),该试剂盒可根据5'-NCR和核心区域的变异区分HCV的1、2、3、4、6基因型以及1a、1b、2a/c、2b、3a、3b、3c、3k、4a、4b、4c/d、4e、4f、4h、5a、6a/b、6g、6f/q、6m、7a亚型。检查了266例IVDU(93.2%为男性;中位年龄:25±6.82岁)和454例非吸毒者(51.3%为男性;中位年龄:56.5±16.06岁)的HCV基因型分布。在IVDU组中,按频率依次观察到基因型1a、3a、1b、4c/d、2b、4、3,在1例患者中检测到基因型1、2a/c和混合基因型(1 + 3a)。在非IVDU组中,按频率依次观察到基因型1b、1a、3a、1、2a/c、4,在1例患者中检测到基因型2b、4c/d、5a、6a/b、6和混合基因型(3 + 4)。基因型1a和3a在IVDU组中显著更高(p < 0.00001,p < 0.00001),而1b在非IVDU患者中显著更高(p < 0.00001)。基因型1a和3a在年轻男性中更常见(p < 0.00001,p = 0.000163),而1b在中年女性中更高(p < 0.00001)。外国国民中基因型1b(p = 0.021)和3a(p = 0.012)的发生率高于土耳其患者。当按年份检查HCV基因型分布时,观察到基因型1b和1a的百分比在下降,而基因型3a的百分比在上升。结果,在本研究中,观察到IVDU中的HCV基因型分布与非IVDU的普通人群不同。发现基因型1a和3a在IVDU组中更常见。与我国其他地区一样,普通人群中基因型1b最常见。与过去几年相比,基因型3a显著增加。在我们的研究中,世界各地存在的基因型的确定可能归因于居住在我们城市的外国国民,并且我们所在地区是一个旅游中心。还需要调查多年来IVDU是否有所增加。