Diagnosis of congenital cytomegalovirus infection by detection of viral DNA in dried blood spots

Adenovirus DNA is detected at increased frequency in Guthrie cards from children who develop acute lymphoblastic leukaemia

“…And, as is the case with children infected post-natally, adenoviral DNA is retained in latently infected T lymphocytes for years following primary infection (Garnett et al, 2002). Thus, the finding of species C adenovirus in neonatal blood spots is evidence of prenatal infection, as is the case for other prenatal viral infections (Barbi et al, 1996;Johansson et al, 1997;Fischler et al, 1999).…”

“…Three uniform discs, 3 mm in diameter, were punched from one of the four blood spots from the stored material and the DNA extracted as previously described with minor modifications (Barbi et al, 1996).…”

A pilot study using residual newborn dried blood spots to assess the potential role of cytomegalovirus and Toxoplasma gondii in the etiology of congenital hydrocephalus

“…Second, previous studies have indicated that neonatal screening for T. gondii IgM using DBS detects ~52-87% of infected newborns (Lebech et al, 1999;Paul et al, 2000;Gilbert et al, 2007). Similarly, the sensitivity of CMV DBS DNA testing approaches 100% when testing symptomatic newborns (Barbi et al, 1996;Snijdewind et al, 2012), but ranges from ~30 to 80% when testing unselected population-based newborns (Soetens et al, 2008;Boppana et al, 2010). While this suggests some case-and control-infants with T. gondii or CMV infection might have been missed with an approach that used residual DBS, the study was most likely to miss those with CMV infection who had a low viral load and thus were at low risk for clinical sequelae related to the infection.…”

“…CMV DNA was extracted from DBS using a modified thermal shock method (Shibata et al, 1994;Barbi et al, 1996). Three 3.2 mm punches were placed in a 2 ml microfuge tube with 60 μl of Glutamine-free Minimum Essential Medium and soaked for 2 hr at room temperature, then incubated at 55 °C for 1 hr, then 100 °C for 7 min, and then transferred to ice for rapid cooling.…”

Prevalence of Congenital Cytomegalovirus Infection Assessed Through Viral Genome Detection in Dried Blood Spots in Children with Autism Spectrum Disorders

“…A 3-mm diameter circle punched out of the DBS was used for CMV-DNA extraction, and nucleic acid was amplified as previously described (64) by an operator blinded to the characteristics of the cases. Stringent control measures were applied to prevent both carryover and contamination (65). In brief, DNA was extracted from the circle of DBS by adding 35 μl of cell culture medium (minimum essential medium) followed by thermal shock (55˚C for 60 min and 100˚C for 7 min, centrifugation at 11,200 rcf, and the supernatant was frozen at −80˚C overnight) and amplified using a nested polymerase chain reaction (PCR) designed to amplify one region in the GP58 gene (66).…”