Rouse DJ, Fette LM, Hughes BL, Saade GR, Dinsmoor MJ, Reddy UM, et al. Noninvasive prediction of congenital cytomegalovirus infection after maternal primary infection. Obstet Gynecol 2022;139.
The authors provided this information as a supplement to their article.
©2022 American College of Obstetricians and Gynecologists. Page 1 of 6
Appendix 1. Trial Sites and Study Personnel
Each clinical center consisted of one or more additional performance sites as listed below. Also listed are members of the Eunice Kennedy Shriver National Institute of Child Health andHuman Development Maternal-Fetal Medicine Units Network who contributed to the research in addition to the authors.
Clinical Centers:
Brown University, Providence, RI – B. Hughes, D. Rouse, D. Allard, E. Werner, J. Rousseau, L. Beati, J.
Milano, J. Lee
University of Texas Medical Branch, Galveston, TX – G. Saade, A. Salazar, L. Pacheco, J. Patel, D. Carlson, K. Smith, A. Nounes, J. DeVolder
Northwestern University, Chicago, IL – G.Mallett, W.Grobman, A. Peaceman NorthShore University Health System, Evanston, IL – M. Dinsmoor, K. Paycheck
Columbia University, New York, NY – C. Gyamfi-Bannerman, S. Bousleiman, R. Wapner V. Carmona, M.
Talucci
Christiana Care, Wilmington, DE – M. Hoffman, A. Vanneman
St. Peter’s University Hospital, New Brunswick, NJ – K. Palomares, C. Perez Drexel University, Philadelphia, PA – L. Plante, C. Tocci,
New York Presbyterian Queens, Flushing, NY – D. Skupski R. Chan-Akeley Lehigh Valley Hospital, Allentown, PA
University of Utah Health Sciences Center, Salt Lake City, UT – M. Varner, K. Hill, A. Sowles LDS Hospital, Salt Lake City, UT – C. Meadows
McKay-Dee Hospital, Ogden, UT – S. Dellerman
Intermountain Med Center, Salt Lake City, UT – L. Hansen, S. Esplin
University of North Carolina at Chapel Hill, Chapel Hill, NC – W. Goodnight, K. Clark, J. Thorp, S. Timlin WakeMed Health & Hospitals, Raleigh, NC – C. Beamon, H. Byers
Prisma Health, Greenville, SC – K. Eichelberger, A. Moore
University of Alabama at Birmingham, Birmingham, AL – A. Tita, S. Harris, L. Harper, J. Biggio, M. Parks, J. Sheppard
The Ohio State University, Columbus, OH– M. Costantine, A. Bartholomew, M. Landon, J. Iams, C.
Shellhaas, K. Markham, B. Rink, C. Buhimschi, F. Johnson, L. Webb
Wright State University Miami Valley Hospital, Dayton, OH – D. McKenna, K. Fennig, K. Snow
Rouse DJ, Fette LM, Hughes BL, Saade GR, Dinsmoor MJ, Reddy UM, et al. Noninvasive prediction of congenital cytomegalovirus infection after maternal primary infection. Obstet Gynecol 2022;139.
The authors provided this information as a supplement to their article.
©2022 American College of Obstetricians and Gynecologists. Page 2 of 6
Duke University, Durham, NC – G. Swamy, T. Bishop, J. Ferrara
University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, CO – R. Gibbs, K. Hale, K.D.
Heyborne, J. Phipers
MetroHealth Medical Center-Case Western Reserve University, Cleveland, OH – E. Chien, W. Dalton University Hospitals, Cleveland, OH – D. Hackney, A. Mayle
UT Health- University of Texas Health Science Center at Houston, Children's Memorial Hermann Hospital, Houston, TX – S. Chauhan, F. Ortiz, B. Sibai
Stanford University, Stanford, CA – Y. El-Sayed, C. Willson, N. Aziz, D. Lyell, A. Girsen, K. Sherwi
University of Texas Southwestern Medical Center, Dallas, TX – B. Casey, L. Moseley, J. Price, T. Thomas, L.
Fay-Randall, A. Sias, M. Garcia
University of Pennsylvania, Philadelphia, PA – S. Parry, J. Craig
University of Pittsburgh, Pittsburgh, PA – H. Simhan, M. Bickus, F. Facco, M. Birsic
Madigan Army Medical Center, Joint Base Lewis-McChord, Tacoma, WA – P. Napolitano, L. Imbruglio, E.
Hemman, J. Pates, L. Foglia
Data Coordinating Center: The George Washington University Biostatistics Center, Washington, DC – E.
Thom, R. Clifton, L. Fete, L. Mele, V. L. Flowers-Fanomezantsoa, T. Boekhoudt
Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD – U.
Reddy, C. Spong, S. Pagliaro
Rouse DJ, Fette LM, Hughes BL, Saade GR, Dinsmoor MJ, Reddy UM, et al. Noninvasive prediction of congenital cytomegalovirus infection after maternal primary infection. Obstet Gynecol 2022;139.
The authors provided this information as a supplement to their article.
©2022 American College of Obstetricians and Gynecologists. Page 3 of 6
Three-variable model
Appendix 2. Prediction Model of Congenital Cytomegalovirus Infection
Factors Adjusted OR (95% CI)
IgG avidity <32% (low) 3.09 (1.60 – 5.96)
IgM antibody index ≥4.5 (high) 3.90 (2.15 – 7.07)
Virus (detectable) 1.86 (1.04 – 3.32)
Goodness of fit: Hosmer and Lemeshow test p-value=0.24
Logistic regression equation for prediction of congenital CMV infection
Predicted probability of congenital CMV infection = exp(w) ÷ [1+exp(w)], where w= - 2.8966 + 1.1275(low IgG avidity) + 1.3597 (high IgM) + 0.6185 (viral load detectable)
Rouse DJ, Fette LM, Hughes BL, Saade GR, Dinsmoor MJ, Reddy UM, et al. Noninvasive prediction of congenital cytomegalovirus infection after maternal primary infection. Obstet Gynecol 2022;139.
The authors provided this information as a supplement to their article.
©2022 American College of Obstetricians and Gynecologists. Page 4 of 6
Appendix 3. Predicted Risk of Congenital Cytomegalovirus Infection Based on Individual Maternal Characteristics
Maternal Characteristics Predicted %
probability
(95% CI)
low IgG avidity, high IgM index, virus detectable 55.2 (42.0 – 67.7) low IgG avidity, low IgM index, virus detectable 24.0 (15.3 – 35.6) low IgG avidity, low IgM index, virus not detectable 14.6 (8.9 – 22.9) high IgG avidity, low IgM index, virus not detectable 5.2 (2.8 – 9.4)
Rouse DJ, Fette LM, Hughes BL, Saade GR, Dinsmoor MJ, Reddy UM, et al. Noninvasive prediction of congenital cytomegalovirus infection after maternal primary infection. Obstet Gynecol 2022;139.
The authors provided this information as a supplement to their article.
©2022 American College of Obstetricians and Gynecologists. Page 5 of 6
Appendix 4. Receiver operating curve (ROC) for the prediction of congenital cytomegalovirus infection (three variable model).
Rouse DJ, Fette LM, Hughes BL, Saade GR, Dinsmoor MJ, Reddy UM, et al. Noninvasive prediction of congenital cytomegalovirus infection after maternal primary infection. Obstet Gynecol 2022;139.
The authors provided this information as a supplement to their article.
©2022 American College of Obstetricians and Gynecologists. Page 6 of 6
Appendix 5. Serum Quantitative PCR
Viral DNA was extracted using High Pure Viral Nucleic Acid Extraction Kit (Roche) from 200 ul of plasma with a final elution volume of 50 ul. For the PCR reaction 5ul of sample was added to duplicate wells containing 25 uL mixture of 15 uL of SybrSelect (Invitrogen), 300 nM of primers (Integrated DNA Technologies) and water. Primers were designed to the immediate early 1 (IE1) gene as previously described.1 qPCR conditions consisted of a 2 minute cycle at 50°C, followed by 10 minutes at 95°C, 40 cycles of denaturation at 95°C for 15 seconds and combined annealing/extension at 63°C for 1 minute.
Serial dilutions of pre-quantitated AD169 virus from advanced biotechnologies with a range of 1-105 copies per mL were ran in triplicate to create a standard curve. Virus was considered detected at >100 copies per mL. The sample was considered positive for CMV by qPCR if at least 2 out of 6 replicates had virus detected. Samples were repeated until 2 replicates were positive or 6 replicates were completed, whichever came first.
REFERENCE
1. Kalpoe JS, Kroes AC, de Jong MD, Schinkel J, de Brouwer CS, Beersma MF, Claas EC. Validation of clinical application of cytomegalovirus plasma DNA load measurement and definition of
treatment criteria by analysis of correlation to antigen detection. J Clin Microbiol. 2004
Apr;42(4):1498-504. doi: 10.1128/JCM.42.4.1498-1504.2004. Erratum in: J Clin Microbiol. 2004 Oct;42(10):4917. PMID: 15070995; PMCID: PMC387533.
