title: Dried-blood sampling for epstein-barr virus immunoglobulin G (IgG) and IgA serology in nasopharyngeal carcinoma screening.
abstract: Dried-blood (DB) samples on filter paper are considered clinical specimens for diagnostic use because of the ease of collection, storage, and transport. We recently developed a synthetic-peptide-based immunoglobulin A (IgA) (EBNA1 plus viral capsid antigen [VCA]-p18) enzyme-linked immunosorbent assay (ELISA) for nasopharyngeal carcinoma (NPC) screening. Here, we evaluate the use of two filter papers for DB sampling, i.e., Schleicher & Schuell (S&S) no. 903 and Whatman no. 3; the DB samples were either taken directly from a finger prick or spotted from a Vacutainer blood collector. The elution of DB samples on filter paper was optimized and tested for IgG and IgA reactivity by ELISA (EBNA1 plus VCA-p18) and compared to simultaneously collected plasma samples. The results showed that both types of filter paper can be used for sample collection in NPC diagnosis by using either finger prick or blood spot sampling. Both DB sampling methods produced comparable ELISA (EBNA1 plus VCA-p18) results for IgG and IgA reactivity in 1:100-diluted plasma samples. DB samples of whole blood or finger prick blood show correlation coefficients (r(2)) of 0.825 to 0.954 for IgA on S&S no. 903 filter paper, 0.9133 to 0.946 for IgA on Whatman no. 3 filter paper, 0.807 to 0.886 for IgG on S&S no. 903 filter paper, and 0.819 to 0.934 for IgG on Whatman no. 3 filter paper. Using plasma IgA as a reference, DB sampling showed sensitivities and specificities of 75.0 to 96.0% and 93.5 to 100%, respectively. DB samples could be stored at 37 degrees C for 1 to 4 weeks on S&S no. 903 filter paper and 1 to 6 weeks on Whatman no. 3 filter paper without a significant loss of reactivity, with provision of transport options for tropical conditions. IgA proved to be more stable than IgG. Whatman no. 3 filter paper is a more economical yet diagnostically comparable alternative to S&S no. 903 filter paper. Finger prick DB sampling is proposed for NPC diagnosis, particularly for remote hospitals and field screening studies.
authors.affiliation: Department of Pathology, Vrije Universiteit Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Fachiroh
authors.forename: P R
authors.initials: PR
authors.lastname: Prasetyanti
authors.forename: D K
authors.initials: DK
authors.lastname: Paramita
authors.forename: A T
authors.initials: AT
authors.lastname: Prasetyawati
authors.forename: D W
authors.initials: DW
authors.lastname: Anggrahini
authors.forename: S M
authors.initials: SM
authors.lastname: Haryana
authors.forename: J M
authors.initials: JM
authors.lastname: Middeldorp
mesh_terms: D000956:Antigens, Viral / Q000276:immunology; D001800:Blood Specimen Collection / Q000379:methods*; D036022:Capsid Proteins / Q000276:immunology; D002277:Carcinoma / Q000175:diagnosis / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis / Q000821:virology; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood*; D007074:Immunoglobulin G / Q000097:blood*; D008403:Mass Screening; D009303:Nasopharyngeal Neoplasms / Q000175:diagnosis / Q000821:virology; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; C041788:Epstein-Barr viral capsid antigen; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D010455:Peptides; C417029:EBV-encoded nuclear antigen 1