Parker, Simon Philip; (2002) The development and evaluation of dried blood spot technology for the study of blood borne infections. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

The use of capillary whole blood, blotted onto filter paper was introduced by Guthrie in 1963 as an ideal means of screening for metabolic diseases in neonates. These 'dried blood spot samples' (DBS) contain IgG antibodies that are primarily of maternal origin and thus provide an ideal sample for prevalence studies in women. Studies using simulated DBS, showed that antibody could be retrieved from DBS adequately stored for long periods of time and that the elution of antibody from the filter paper matrix was reliable. It was then possible to develop and modify low cost immunoassays for the detection of antibodies to hepatitis C virus (HCV), human T cell lymphotropic virus (HTLV) and HIV-1. This was followed by the installation and maintenance of extensive anonymous epidemiological surveys using DBS taken from the Guthrie cards of neonates born in south-east England. The anti-HIV-1 and HTLV screening assays were based on a modification of commercial agglutination assays whilst an 'in house' ELISA was developed for the detection of anti-HCV IgG. The presence of specific antibody was confirmed using selected commercial assays and an 'in house' anti-HIV-1 serotyping ELISA adapted for use with DBS. Over a 15 month period, more than 120,000 DBS were collected and tested using the three serological algorithms. Screening and confirmation was possible using a single eluate derived from two DBS punched from each Guthrie card. Epidemiological data, including mother's age and country of origin, was made available from a proportion of the DBS tested. Results confirmed a higher prevalence of HCV in women residing in Inner London and those born in southern Europe or Asia. Other results confirmed that the highest prevalence of HIV-1 was in those women residing in Inner London and born in sub-Saharan Africa and for HTLV-I in women born in the Caribbean. Low cost, long-range postal and shipping surveys for anti-HCV, HTLV and HIV-1 were implemented in South Africa, and Pakistan, countries with wide ranging epidemiological needs. The serological algorithms described and the renewed interests in the application of DBS for epidemiological study have led to studies in many other countries.

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