Pp65 antigenemia, plasma real-time PCR and DBS test in symptomatic and asymptomatic cytomegalovirus congenitally infected newborns.

BACKGROUND: Many congenitally cytomegalovirus-infected (cCMV) neonates are at risk for severe consequences, even if they are asymptomatic at birth. The assessment of the viral load in neonatal blood could help in identifying the babies at risk of sequelae. METHODS: In the present study, we elaborated the results obtained on blood samples collected in the first two weeks of life from 22 symptomatic and 48 asymptomatic newborns with cCMV diagnosed through urine testing. We evaluated the performances of two quantitative methods (pp65 antigenemia test and plasma Real-time PCR) and the semi-quantitative results of dried blood sample (DBS) test in the aim of identifying a valid method for measuring viral load. RESULTS: Plasma qPCR and DBS tests were positive in 100% of cases, antigenemia in 81%. Only the latter test gave quantitatively different results in symptomatic versus asymptomatic children. qPCR values of 103 copies/ml were found in 52% of newborn. "Strong" DBS test positivity cases had higher median values of both pp65 positive PBL and DNA copies/ml than cases with a "weak" positivity. CONCLUSIONS: As expected antigenemia test was less sensitive than molecular tests and DBS test performed better on samples with higher rates of pp65 positive PBL and higher numbers of DNA copies/ml. The prognostic significance of the results of these tests will be evaluated on completion of the ongoing collection of follow-up data of these children.

Modification of CMV DNA detection from dried blood spots for diagnosing congenital CMV infection.

BACKGROUND: Detection of viral DNA in dried blood spots using the Guthrie card (DBS test) is a reliable and practical method of diagnosing congenital cytomegalovirus (CMV) infection. The test lends itself to epidemiological studies to establish the prevalence of the infection, but also to neonatal screening for secondary prevention of sequelae. These applications would be facilitated if it were possible to use smaller samples and do the test on pools of individual cases. OBJECTIVE: To ascertain whether doing the test on smaller, pooled samples still accurately identifies neonates with congenital CMV infection. STUDY DESIGN: We tested DBS from: (A) 39 laboratory reference cases; (B) 156 neonates suspected of having congenital CMV infection; (C) 119 children examined for the retrospective diagnosis of congenital CMV; (D) mock specimens prepared with known amounts of viral DNA. RESULTS: The test using only one third of the usual amount of dried blood was 100% sensitive and specific compared to the standard DBS test (A) and to viral isolation (A and B). Pools of three single cases gave the same results as viral isolation (B) and the small-sample test (B and C). All the versions of the test gave a detection limit of 400 copies/ml. CONCLUSIONS: The modified procedure can accurately diagnose congenital CMV infection. It achieves savings in both the patient material and the costs of testing.

Prevalence of anti-poliovirus type 1, 2 and 3 antibodies in unvaccinated Italian travelers.

BACKGROUND: Immunization against poliomyelitis is recommended for international travelers to developing countries. However, the level of antibodies varies even in previously unvaccinated persons, due to wild-type or vaccine-type infections in the eldest travelers. METHODS: In 1999, we conducted a seroprevalence study in the Lombardy region (northern Italy), using sera collected in 1994 from a population aged 50 to 59 years. The study subjects were consecutive, randomly selected travelers enrolled in an anti-hepatitis A virus antibody study. Neutralizing antibodies were titrated on Vero cells in microtiter plates. Each serum dilution (1:8 to 1:256) was challenged against 100 tissue culture infective doses of the three Sabin strains. Titers> 1:8 were considered to be protective. RESULTS: We studied 98 travelers, 59 male and 39 female, of mean age 54 years. Seventy-three (74.4%) reported previous travel abroad, but none had been vaccinated against polio. Dietary habits included consumption of seafood in 74.4% and raw vegetables from their own garden in 52.1%. The seroprevalences for neutralizing antibodies against poliovirus type 1, type 2 and type 3 were 86.7%, 89.9%, and 86.7%, respectively. All travelers presented protective antibody titers against at least one of the three viral types. Protective antibody titers were unrelated to travel history or dietary habit. CONCLUSIONS: A high proportion of the previously unvaccinated adults in our sample presented protective immunity to polioviruses. This observation may have implications for cost-effectiveness analysis of generalized polio vaccination in adult Italian travelers.