As part of the CMV and Hearing Multicenter Screening (CHIMES) research

As part of the CMV and Hearing Multicenter Screening (CHIMES) research 72 239 newborns were screened for CMV by fast culture and real-time PCR of saliva samples. Around 10-15% of the asymptomatic babies and about 50% of babies with medical abnormalities at delivery (“symptomatic” cCMV) will establish sequelae including SNHL beyond the neonatal period 4 5 In a big multi-center research (CMV and Hearing Multicenter Testing or CHIMES Research) we proven that testing newborns for Varenicline CMV disease by real-time PCR tests of saliva examples for CMV DNA was extremely sensitive and particular 6. The aim of this research was to determine if the real-time PCR assay identified more newborns with CMV than rapid culture of saliva specimens the Varenicline latter being the gold standard for identifying CMV-infected newborns. Materials and Methods Study Design Infants born at seven hospitals in the United States between June 2008 and March 2012 were enrolled prospectively in the National Institute on Deafness and other Communication Disorders (NIDCD) sponsored CHIMES study6. Between June 2008 and December 2009 35 334 newborns were screened for CMV by rapid culture and real-time PCR performed on saliva swabs placed in transport medium (liquid saliva specimens) as described previously6. From January 2010 to March 2012 36 905 newborns were screened for CMV by PCR of dried saliva specimens followed by rapid culture of PCR-positive specimens. The CMV testing protocols for PCR and rapid culture have been described 6. Positive screening results (PCR or rapid culture of newborn saliva) were confirmed by follow-up rapid culture and PCR testing of urine and saliva samples obtained within 3-6 weeks of birth. The concordance between PCR and rapid culture of newborn screening saliva samples was compared to determine if use of saliva PCR for screening identifies more infants with congenital CMV infection. Determination of viral load Viral load values expressed as international units (IU/mL) based on calibration to the CMV WHO Standards 7 and the number of fluorescent cells on rapid culture were likened in discordant examples to see whether the discordance between real-time PCR and fast culture was supplementary to low viral fill. Statistical evaluation As the real-time PCR and fast culture are combined examples the McNemar’s check was utilized to evaluate the outcomes of both assays. Viral fill between discordant and concordant samples was analyzed using unpaired t check. Results Outcomes of newborn CMV testing (Shape 1) Shape 1 Algorithm for CMV PCR and Quick Culture Testing Lamb2 of Infants Shape 1 Algorithm for CMV PCR and Quick Culture Testing of Infants From the 73 239 babies screened for CMV through the research period 284 (0.4%) babies tested positive by PCR or quick tradition of saliva and were enrolled for verification of cCMV. The mean (±SD) period between test collection and efficiency of PCR and fast tradition was 9.2±5.3 times and 9.1±5.5 times respectively (p=1). On confirmatory tests 18 babies had Varenicline adverse PCR and fast tradition of saliva and urine examples and were thought to possess false positive testing results. Testing PCR and fast culture assay outcomes for the 266 babies confirmed to have cCMV were compared. Of these newborn saliva samples from 252 (94.7%) were positive by both PCR and rapid culture. Discordant results between PCR and rapid culture were observed in fourteen infants and of these 13 were PCR positive and rapid culture negative whereas one was rapid culture positive and PCR negative. The number of samples with discordant results between the two screening assays was significantly higher when tested by rapid culture than PCR (McNemar’s test; p = 0.003). Duration between collection of screening samples and testing When compared to samples with concordant results the mean duration (± SD) from collection to testing was longer for samples with discordant results by both PCR (9.5±5.3 vs 12.4±3.3 days; p=0.05) and rapid culture (8.8±5.4 vs 14.6±5 days; p=0.02) Varenicline respectively. Among samples with discordant results there was no significant difference in duration between sample collection to performance of PCR or rapid culture (p=1). Viral load in discordant specimens The median viral load was not significantly different between discordant (1.86×105 IU/ml range: 6.4×102 to 4.8×107 IU/mL) and concordant samples 2.5 IU/ml range: 1×103 to 3×1010 IU/ml p=0.7). The one sample that was rapid culture positive and PCR negative had only four positive fluorescent cells per.