Usefulness of blood and urine samples collected on filter paper in detecting cytomegalovirus by the polymerase chain reaction technique

• Published in: Journal of virological methods 2001-09, Vol.97 (1), p.159-164
• Main Authors: Yamamoto, A.Y., Mussi-Pinhata, M.M., Pinto, P.C.G., Figueiredo, L.T.M., Jorge, S.M.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Blood Specimen Collection - methods; Cell Line, Transformed; Congenital cytomegalovirus infection; Cytomegalovirus - genetics; Cytomegalovirus - isolation & purification; Cytomegalovirus Infections - blood; Cytomegalovirus Infections - urine; Cytomegalovirus Infections - virology; DNA amplification; DNA, Viral - analysis; DNA, Viral - genetics; Dried spot blood; Dried spot urine; Filtration; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Human cytomegalovirus; Human viral diseases; Humans; Infant; Infectious diseases; Medical sciences; Microbiology; Micropore Filters - virology; Molecular immunology; Multiplex PCR; Neonatal diagnosis; Nested PCR; Polymerase Chain Reaction; Sensitivity and Specificity; Techniques; Viral diseases; Viral diseases of the lymphoid tissue and the blood. Aids
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(01)00347-0

A rapid test for the diagnosis of congenital CMV infection is still needed. This study evaluated the usefulness of dried blood and urine samples collected on filter paper for detecting cytomegalovirus (CMV) by the polymerase chain reaction (PCR) assay compared with the use of liquid urine. Samples were obtained from 332 infants aged 1–7 days. Liquid urine samples were collected into bags, cultured in human fibroblasts, and processed using a multiplex PCR technique. Dried urine samples were obtained by placing a piece of filter paper in contact with the infant's genitals. The heels of neonates were punctured and capillary blood was blotted onto filter paper and dried. Dried blood and urine specimens were analyzed by multiplex PCR and nested-PCR assays. A diagnosis of congenital CMV infection was established by isolating the virus, and by detecting viral DNA in the liquid urine. Of the 332 liquid urine samples collected from 332 neonates, seven (2.1%) were positive for CMV and 325 were negative, by both cell culture and PCR assay. In dried samples, CMV DNA was detectable only with a nested PCR assay. Compared with known CMV infection status, 5/7 (71.4%) neonates were positive for congenital CMV infection using dried blood samples. All 325 uninfected neonates were negative. In the dried urine samples, 4/4 CMV-infected infants gave positive tests, and all 262 uninfected infants were negative. Although further improvements in sample collection and/or processing are still needed, PCR testing on dried urine or blood collected on filter paper is a promising approach in the diagnosis of neonatal CMV infection.