Dried blood spot as alternative specimen for molecular epidemiology studies among HCV/HIV coinfected patients

IF 3 4区 医学 Q2 INFECTIOUS DISEASES
Geane Flores , Barbara Vieira Lago , Amanda R Caetano , Jessica Silva , Vanessa Marques , Carlos Eduardo Brandão-Mello , Marcia Amendola-Pires , Jose Pilotto , Lia Lewis-Ximenez , Livia Melo Villar
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Abstract

Background

Immunodeficiency Virus (HIV) and Hepatitis C Virus (HCV) share the same routes of transmission, therefore, co-infection by both viruses represents a challenge to the goal of eliminating viral hepatitis as a public health threat. There are an estimated 2.3 million people living with HIV/HCV worldwide. Most of these cases affect vulnerable populations located in places with low infrastructure. Because of this, the use of alternative samples such as Dried Blood on Spot (DBS) would facilitate access to diagnosis and HCV treatment. The aim of this study is to evaluate the HCV genetic variability in HIV/HCV individuals by correlating paired serum and DBS samples.

Methods

A total of 14 HIV/HCV individuals, recruited from reference outpatient clinics in the city of Rio de Janeiro/Brazil, were included. From them, 64 % were man, mean of age 54±7. HCV RNA from both serum and DBS samples was RT-PCR amplified and sequenced with HCV NS5B-specific oligonucleotides. All positive samples were submitted to phylogenetic analysis.

Results

Serum mean HCV load was 6.2 ± 0.5 log IU/mL. All patients presented undetectable HIV RNA. The distribution of HCV genotypes/subgenotypes was 1a (4/14); 1b (5/14); 3a (4/14); and 4d (1/14). Most paired serum and DBS samples showed concordant results (genetic distance: 0.0 to 0.16). One individual showed discordance in the subtypes between serum and DBS. Three individuals presented the 316 N Resistance Associated Mutation (RAS) in both serum and DBS.

Conclusion

Our results demonstrate the applicability of DBS for HCV molecular tracking in HIV/HCV coinfected patients for viral genomic surveillance in key and vulnerable populations.
干血斑作为HCV/HIV合并感染患者分子流行病学研究的备选标本
免疫缺陷病毒(HIV)和丙型肝炎病毒(HCV)具有相同的传播途径,因此,这两种病毒的合并感染对消除病毒性肝炎这一公共卫生威胁的目标构成了挑战。据估计,全世界有230万人感染艾滋病毒/丙型肝炎病毒。这些病例大多影响基础设施薄弱地区的弱势人群。因此,使用现场干血(DBS)等替代样本将有助于获得诊断和HCV治疗。本研究的目的是通过配对血清和DBS样本的相关性来评估HIV/HCV个体的HCV遗传变异性。方法从巴西里约热内卢市参考门诊共招募14名HIV/HCV患者。其中男性占64%,平均年龄54±7岁。对血清和DBS样本中的HCV RNA进行RT-PCR扩增,并用HCV ns5b特异性寡核苷酸测序。所有阳性样本均进行系统发育分析。结果血清平均HCV载量为6.2±0.5 log IU/mL。所有患者的HIV RNA均检测不到。HCV基因型/亚基因型分布为1a (4/14);1 b (5/14);3 (4/14);4d (1/14)大多数配对血清和DBS样品显示一致的结果(遗传距离:0.0 ~ 0.16)。1例患者血清亚型与DBS亚型不一致。3例患者血清和DBS均出现316n耐药相关突变(RAS)。结论DBS用于HCV分子跟踪的方法适用于重点人群和易感人群的HCV病毒基因组监测。
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来源期刊
CiteScore
5.50
自引率
0.00%
发文量
925
审稿时长
41 days
期刊介绍: The Brazilian Journal of Infectious Diseases is the official publication of the Brazilian Society of Infectious Diseases (SBI). It aims to publish relevant articles in the broadest sense on all aspects of microbiology, infectious diseases and immune response to infectious agents. The BJID is a bimonthly publication and one of the most influential journals in its field in Brazil and Latin America with a high impact factor, since its inception it has garnered a growing share of the publishing market.
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