Department of Pediatrics, Shengjing Hospital of China Medical University, Shenyang, China.
School of Forensic Medicine, China Medical University, Shenyang, China.
Front Cell Infect Microbiol. 2024 Jul 19;14:1430302. doi: 10.3389/fcimb.2024.1430302. eCollection 2024.
Human cytomegalovirus (HCMV) is the most common viral infection seen in newborns. The major route of transmission for acquired human cytomegalovirus infection is breast milk from mothers who are HCMV seropositive to the infants. Thus, a rapid, economical, and simple method to perform HCMV test in breast milk is crucial and necessary for preventing acquired HCMV infection, especially in underdeveloped regions with limited laboratory resources.
In this study, an effective technique for the detection of HCMV was constructed by combining multienzyme isothermal rapid amplification (MIRA) and lateral flow chromatography strip (LFD). Primers for the conserved HCMV sequence UL83 were utilized for MIRA-LFD testing.
Our results showed that the entire MIRA reaction could be completed in 12 minutes at 37°C, and LFD outcomes could be observed visibly after 10 minutes. The detection sensitivity of this method reached 50 copy/μl. Samples of breast milk were examined to compare MIRA-LFD and conventional qPCR. The accuracy of MIRA-LFD was 100%.
The straightforward, rapid, economic features of the test can provide the significant advantages for the prevention of breast milk-acquired cytomegalovirus infection, particularly in resource-limited locations with high seroprevalence of cytomegalovirus.
人巨细胞病毒（HCMV）是新生儿最常见的病毒感染。获得性人巨细胞病毒感染的主要传播途径是来自 HCMV 血清阳性母亲的母乳传染给婴儿。因此，对于预防获得性 HCMV 感染，特别是在实验室资源有限的欠发达地区，快速、经济、简单的母乳 HCMV 检测方法至关重要。
本研究通过结合多酶等温快速扩增（MIRA）和侧流层析条带（LFD）构建了一种有效的 HCMV 检测技术。用于 MIRA-LFD 检测的引物是针对保守的 HCMV 序列 UL83 的。
我们的结果表明，整个 MIRA 反应可以在 37°C 下 12 分钟内完成，10 分钟后可以肉眼观察到 LFD 结果。该方法的检测灵敏度达到 50 拷贝/μl。我们检测了母乳样本，比较了 MIRA-LFD 和常规 qPCR。MIRA-LFD 的准确率为 100%。
该检测方法简单、快速、经济，为预防母乳获得性巨细胞病毒感染提供了显著优势，特别是在巨细胞病毒血清阳性率较高、资源有限的地区。
