Evaluation of dried blood spot samples for screening of hepatitis C and human immunodeficiency virus in a real-world setting

Abstract

Both hepatitis C virus (HCV) infection and human immunodeficiency virus (HIV) infection areunderdiagnosed, particularly in low-income countries and in difficult-to-access populations. Our aim was to develop and evaluate a methodology for the detection of HCV and HIV infection based on capillary dry blood spot (DBS) samples taken under real-world conditions. We carried out a crosssectional study of 139 individuals (31 healthy controls, 68 HCV-monoinfected patients, and 40 HCV/ HIV-coinfected patients). ELISA was used for anti-HCV and anti-HIV antibody detection; and SYBR Green RT-PCR was used for HCV-RNA detection. The HIV serological analysis revealed 100% sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). The HCV serological analysis revealed a sensitivity of 92.6%, specificity of 100%, PPV of 100%, and NPV of 79.5%. Finally, the HCV-RNA detection test revealed a detection limit of 5 copies/µl with an efficiency of 100% and sensitivity of 99.1%, specificity of 100%, PPV of 100%, and NPV of 96.9%. In conclusion, our methodology was able to detect both HCV infection and HIV infection from the same DBS sample with good diagnostic performance. Screening for HCV and HIV using DBS might be a key strategy in the implementation of national programs for the control of both infections.Hepatitis C virus (HCV) infection is a major health problem worldwide. Approximately 71 million persons live with HCV infection, and around 80% are undiagnosed 1 . Infection by human immunodeficiency virus (HIV) continues to be a major public health issue, with a global prevalence of 0.8% among adults, of whom around 30% are undiagnosed 2 . Both HIV and HCV share routes of transmission, and their diagnosis is a priority in public health strategies throughout the world 1,2 . On the one hand, HIV infection is diagnosed increasingly frequently in resource-limited settings through implementation of HIV testing programs; however, HIV testing is still not reaching some communities (eg, people who inject drugs [PWID], sex workers, homeless persons), resulting in late presentation 3 . On the other hand, diagnosis of HCV remains problematic for many low-income countries and difficult-to-access populations in developed countries 4 , where most individuals remain unaware of their HCV status until advanced stages of the disease 5-7 .Traditionally, diagnosis of HIV and HCV infection has been based on the detection of antibodies, antigens, and the viral genome in blood samples obtained by routine venous blood collection 5,8 . However, the use of capillary whole blood in dried blood spot (DBS) specimens may be an excellent alternative in both serological and nucleic acid testing (NAT) assays for the screening of HIV and HCV infection, since these approaches are less invasive, samples can be stored and transported without refrigeration, and highly trained personnel are not necessary 5,9,10 . Therefore, DBS may facilitate diagnosis of HIV and HCV infection in settings where access to screening is difficult. In fa...

Screening for Human Immunodeficiency Virus Infection by Use of a Fourth-Generation Antigen/Antibody Assay and Dried Blood Spots: In-Depth Analysis of Sensitivity and Performance Assessment in a Cross-Sectional Study

“…DBS have become a popular method in a variety of micro-blood-sampling techniques in the life sciences sector (2,5,6). They have gained a significant role in the diagnosis of infectious diseases, especially in the HIV/AIDS field, where they have been used either for diagnosis and monitoring at the individual level or to perform surveillance studies at the population level (3,4,19,20). Since the mid-1980s, extraordinary progress in improving the sensitivity of serological assays for HIV infection has been made continuously, and the fourth-generation assays are now recommended for HIV screening (12-15).…”

“…gold standard for HIV screening due to improved detection during the early phase of infection (11)(12)(13)(14)(15). Although several studies have already analyzed the performance of such assays using DBS (16)(17)(18)(19)(20), we wanted to characterize further the sensitivity in the context of an HIV surveillance program among men who have sex with men (MSM), the Prevagay study. Indeed, two main situations may limit the sensitivity in a high-risk population.…”

Dried Blood Spot in Laboratory: Directions and Prospects

“…DBS technology is also well adapted for use with enzyme-linked immunosorbent assay (ELISA) reactions, allowing high selectivity, reproducibility and specificity of protein detection via antigen/antibody interactions. Specifically, it has been reported on repeated occasion that studies have achieved successful isolation of samples from MC surfaces, which were subsequently analyzed by ELISA for the presence of antibodies against Epstein-Barr virus [50], rubella virus [51], dengue virus [52], hepatitis C virus [53] and HIV virus [54].…”

Evaluation of a hepatitis C virus core antigen assay from venepuncture and dried blood spot collected samples: A cohort study

“…While the good performance of HCV core antigen in plasma has been demonstrated, there are many settings for which the collection or transport of plasma to a central facility is not possible. Dried blood spot (DBS) utility as an alternative sample type in HCV serology and molecular testing (NAT) has been well documented . The major advantages of finger‐stick capillary DBS testing are as follows: (a) simplified collection with no need for phlebotomy (through self‐ or assisted‐collection); (b) easy and affordable transportation of samples (no cold chain transport requirement as samples are stable at room temperature (24°C), with HCV RNA and core antigen stable for a year or more allowing shipment through the mail) and (c) multiple sample availability; as up to five spots are collected, which allows for reflex testing if confirmation is required or multi‐analyte testing.…”