Dried blood spot real-time polymerase chain reaction assays to screen newborns for congenital cytomegalovirus infection
Suresh B Boppana, Shannon A Ross, Zdenek Novak, Masako Shimamura, Robert W Tolan Jr, April L Palmer, Amina Ahmed, Marian G Michaels, Pablo J Sánchez, David I Bernstein, William J Britt, Karen B Fowler, National Institute on Deafness and Other Communication Disorders CMV and Hearing Multicenter Screening (CHIMES) Study, Nitin Arora, Amita Bey, Belinda Blackstone, Jennifer Blumenthal, Valisa Brown, Alice Brumbach, Nazma Chowdhury, Steven Febres-Cordero, Monique Jackson, Mirjam Kempf, David Kimberlin, Noelle Le Lievre, Faye McCollister, Emily Mixon, Misty Purser, Julie Woodruff, Edie Cox, Julie Courtney, Nubia Flores, Molly Ricart, Lisa Schneider, Jennifer West, Jena Colaberardino, Noreen Jeffrey, Anne Maracek, Gretchen E Probst, Cheryl Rosenberg, Diane Sabo, Melissa Calderon, Maria Class, Kristina Feja, Marci Schwab, Daniel Choo, Kate Catalanotto, Linda Jamison, Patty Kern, Kurt Schibler, Maureen Sullivan-Mahoney, Stacie Wethington, Kathy Irving, Delia Owens, Suzanne Roark, Mindy Ware, Cathy Boatman, Jessica Esquivel, Gregory L Jackson, Kathy Katz-Gaynor, April Liehr Townsley, Asuncion Mejías, Kristine E Owen, Peter S Roland, Oscar Rosado, Angela G Shoup, David Sosa, Jessica Santoyo, Elizabeth K Stehel, Lizette Torres, Fiker Zeray, Suresh B Boppana, Shannon A Ross, Zdenek Novak, Masako Shimamura, Robert W Tolan Jr, April L Palmer, Amina Ahmed, Marian G Michaels, Pablo J Sánchez, David I Bernstein, William J Britt, Karen B Fowler, National Institute on Deafness and Other Communication Disorders CMV and Hearing Multicenter Screening (CHIMES) Study, Nitin Arora, Amita Bey, Belinda Blackstone, Jennifer Blumenthal, Valisa Brown, Alice Brumbach, Nazma Chowdhury, Steven Febres-Cordero, Monique Jackson, Mirjam Kempf, David Kimberlin, Noelle Le Lievre, Faye McCollister, Emily Mixon, Misty Purser, Julie Woodruff, Edie Cox, Julie Courtney, Nubia Flores, Molly Ricart, Lisa Schneider, Jennifer West, Jena Colaberardino, Noreen Jeffrey, Anne Maracek, Gretchen E Probst, Cheryl Rosenberg, Diane Sabo, Melissa Calderon, Maria Class, Kristina Feja, Marci Schwab, Daniel Choo, Kate Catalanotto, Linda Jamison, Patty Kern, Kurt Schibler, Maureen Sullivan-Mahoney, Stacie Wethington, Kathy Irving, Delia Owens, Suzanne Roark, Mindy Ware, Cathy Boatman, Jessica Esquivel, Gregory L Jackson, Kathy Katz-Gaynor, April Liehr Townsley, Asuncion Mejías, Kristine E Owen, Peter S Roland, Oscar Rosado, Angela G Shoup, David Sosa, Jessica Santoyo, Elizabeth K Stehel, Lizette Torres, Fiker Zeray
Abstract
Context: Reliable methods to screen newborns for congenital cytomegalovirus (CMV) infection are needed for identification of infants at increased risk of hearing loss. Since dried blood spots (DBS) are routinely collected for metabolic screening from all newborns in the United States, there has been interest in using DBS polymerase chain reaction (PCR)-based methods for newborn CMV screening.
Objective: To determine the diagnostic accuracy of DBS real-time PCR assays for newborn CMV screening.
Design, setting, and participants: Between March 2007 and May 2008, infants born at 7 US medical centers had saliva specimens tested by rapid culture for early antigen fluorescent foci. Results of saliva rapid culture were compared with a single-primer (March 2007-December 2007) and a 2-primer DBS real-time PCR (January 2008-May 2008). Infants whose specimens screened positive on rapid culture or PCR had congenital infection confirmed by the reference standard method with rapid culture testing on saliva or urine.
Main outcome measures: Sensitivity, specificity, and positive and negative likelihood ratios (LRs) of single-primer and 2-primer DBS real-time PCR assays for identifying infants with confirmed congenital CMV infection.
Results: Congenital CMV infection was confirmed in 92 of 20,448 (0.45%; 95% confidence interval [CI], 0.36%-0.55%) infants. Ninety-one of 92 infants had positive results on saliva rapid culture. Of the 11,422 infants screened using the single-primer DBS PCR, 17 of 60 (28%) infants had positive results with this assay, whereas, among the 9026 infants screened using the 2-primer DBS PCR, 11 of 32 (34%) screened positive. The single-primer DBS PCR identified congenital CMV infection with a sensitivity of 28.3% (95% CI, 17.4%-41.4%), specificity of 99.9% (95% CI, 99.9%-100%), positive LR of 803.7 (95% CI, 278.7-2317.9), and negative LR of 0.7 (95% CI, 0.6-0.8). The positive and negative predictive values of the single-primer DBS PCR were 80.9% (95% CI, 58.1%-94.5%) and 99.6% (95% CI, 99.5%-99.7%), respectively. The 2-primer DBS PCR assay identified infants with congenital CMV infection with a sensitivity of 34.4% (95% CI, 18.6%-53.2%), specificity of 99.9% (95% CI, 99.9%-100.0%), positive LR of 3088.9 (95% CI, 410.8-23 226.7), and negative LR of 0.7 (95% CI, 0.5-0.8). The positive and negative predictive values of the 2-primer DBS PCR were 91.7% (95% CI, 61.5%-99.8%) and 99.8% (95% CI, 99.6%-99.9%), respectively.
Conclusion: Among newborns, CMV testing with DBS real-time PCR compared with saliva rapid culture had low sensitivity, limiting its value as a screening test.
Conflict of interest statement
Financial Disclosures: None of the authors had potential conflicts of interest, including specific financial interests and relationships and affiliations (other than those listed in the title page of the manuscript) relevant to the subject of the manuscript.
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Source: PubMed