Abstract
ABSTRACT Introduction: For conducting field studies on human immunodeficiency virus (HIV) serodiagnosis, the use of samples collected on filter paper [dried blood spot (DBS)] is profitable owing to the simplicity in handling and delivering them. Because of these characteristics, the use of DBS is recommended for studies in which the goal is to increase the population access to diagnostic testing, including HIV infection screening. For HIV diagnosis, the conventional strategy firstly uses an enzyme immunoassay as screening test, and the positive sample is confirmed on a complementary assay, as western blotting (WB). Objective: This study aimed at evaluating the analytical performance of WB assay for analyzing DSB samples. Method: One hundred eighteen blood samples collected in filter paper were analyzed by a modified WB. These samples derived from the SampaCentro Study, and they were HIV-antibody-positive in a screening test. In order to assess the reliability of these results, the assay performance was previously certified by employing the sample panel (reference panel) of Instituto Adolfo Lutz (IAL), which consisted of whole blood and plasma paired samples. Results: All of the DBS samples [100% (118/118)] showed the band pattern compatible with the criterion for HIV positivity established by the Ministry of Health. To validate the WB testing, the reciprocal relationship between antibody reactivity of DBS samples and of the respective plasma samples was investigated. Conclusion: WB optimization allowed the achievement of accurate results, and the easy DBS sample collection, transportation and storage will support the use of this immunodiagnostic technique for conducting HIV/acquired immunodeficiency syndrome (Aids) epidemiological surveys.
Highlights
For conducting field studies on human immunodeficiency virus (HIV) serodiagnosis, the use of samples collected on filter paper [dried blood spot (DBS)] is profitable owing to the simplicity in handling and delivering them
western blotting (WB) optimization allowed the achievement of accurate results, and the easy dried blood spots (DBS) sample collection, transportation and storage will support the use of this immunodiagnostic technique for conducting HIV/acquired immunodeficiency syndrome (Aids) epidemiological surveys
Several types of samples may be used in the laboratory diagnosis of HIV infection, such as plasma, serum, whole blood, oral fluid, and blood samples collected on filter paper filter [dried blood spots (DBS)] and saturating it
Summary
It is essential for the world health community to find ways to simplify and improve the accuracy of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (Aids) diagnosis, without reducing the quality of care provided to patients[1]. DBS samples have become widely employed in the serological diagnosis of several illnesses because of facility in transportation, storage, and handling, as well as in seroepidemiologic studies of HIV in remote areas[6,7,8]. The use of these samples minimizes the risks associated to biological transportation and occupational accidents with potentially infected materials[2]. In Brazil, procedures for the laboratory diagnosis of HIV infection are standardized by means of an ordinance by the Ministry of Health, which rules from the type of biologic material to be analyzed to the release of result reports[15]. It must be taken into consideration that reliability of laboratory diagnosis depends on the pre-analytical phase, that is, collection, storage, time and transportation conditions of the sample to the laboratory[17, 19]
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