Hernandez-Alvarado Nelmary, Bierle Craig J, Schleiss Mark R
Department of Pediatrics, Division of Pediatric Infectious Diseases, University of Minnesota Medical School, Minneapolis, MN 55455, USA.
Int J Neonatal Screen. 2023 Dec 20;10(1):1. doi: 10.3390/ijns10010001.
Congenital cytomegalovirus (cCMV) infection is a leading cause of sensorineural hearing loss (SNHL) and neurodevelopmental disabilities in children worldwide. Some regions in the United States and Canada have implemented universal newborn screening for cCMV, which requires molecular diagnostic technologies for identifying cCMV, such as PCR testing of newborn dried blood spots (DBS). This study aimed to evaluate the sensitivity of droplet digital PCR (ddPCR) compared to quantitative real-time PCR to detect CMV DNA in newborn DBS. The limit of detection of various ddPCR primer/probe combinations (singleplex UL55-HEX, singleplex UL83-FAM, and multiplex UL55-HEX/UL83-FAM) was evaluated using the National Institute of Standards and Technology's (NIST) CMV quantitative standard. Singleplex UL55-HEX ddPCR exhibited the lowest limit of detection among the primer/probe combinations tested for ddPCR. UL55 ddPCR was then compared to real-time PCR in 49 infants with confirmed cCMV identified through newborn screening for CMV in saliva swabs and confirmed by a urine test. The results showed that ddPCR was only positive for 59% (29 out of 49) of the cCMV infants, while real-time PCR was positive for 80% (39 out of 49). Due to its lower sensitivity and throughput, ddPCR may not be suitable for cCMV newborn screening.
先天性巨细胞病毒（cCMV）感染是全球儿童感音神经性听力损失（SNHL）和神经发育障碍的主要原因。美国和加拿大的一些地区已实施针对cCMV的新生儿普遍筛查，这需要用于鉴定cCMV的分子诊断技术，例如对新生儿干血斑（DBS）进行PCR检测。本研究旨在评估与定量实时PCR相比，液滴数字PCR（ddPCR）检测新生儿DBS中CMV DNA的灵敏度。使用美国国家标准与技术研究院（NIST）的CMV定量标准评估了各种ddPCR引物/探针组合（单重UL55-HEX、单重UL83-FAM和多重UL55-HEX/UL83-FAM）的检测限。在测试的ddPCR引物/探针组合中，单重UL55-HEX ddPCR的检测限最低。然后，将UL55 ddPCR与实时PCR在49例通过唾液拭子CMV新生儿筛查确诊为cCMV并经尿液检测确认的婴儿中进行比较。结果显示，ddPCR仅在59%（49例中的29例）的cCMV婴儿中呈阳性，而实时PCR在80%（49例中的39例）的婴儿中呈阳性。由于其较低的灵敏度和通量，ddPCR可能不适用于cCMV新生儿筛查。
