Coulibaly T A, Koné A, Sissoko H, Goita A, Cissoko Y, Maiga M, Maiga A I, Kampo M I, Diakité M, Doumbia S, McFall S, Fofana D B
University Clinical Research Center (UCRC).
Université des sciences, des Techniques et des Technologies de Bamako (USTTB).
Rev Mali Infect Microbiol. 2024;19(1):50-55. Epub 2024 Mar 29.
Mother-to-child transmission (MTCT) of hepatitis B virus (HBV) is one of the main causes of chronic hepatitis B in endemic regions such as West Africa. Its prevention constitutes an essential element to eliminate HBV. Without intervention, rates of vertical transmission of HBV vary depending on the level of viral replication. The management of this infection is a major concern, particularly the availability of the viral load at an affordable cost in countries with limited resources such as Mali. This study aimed to develop and validate a method for detecting and quantifying HBV DNA using qPCR in pregnant women, a population at risk of transmitting the virus to newborns.
We enrolled 74 pregnant women with positive AgHBs in this study. Their viral loads were previously determined at the Reference Centre Lab. We designed specific probes and primers for the HBV PreC gene, for detection and quantification by qPCR. We adapted this new qPCR for quantifying HBV DNA on different real-time PCR machines.
Nine out of nine (9/9), 100% samples with a viral load (VL) between 10000-100000 IU/mL and 4/4,100% with a VL > 100,000 IU/mL were detected and quantified. Of the fifty-five (55) samples with a CV of 12-10000 IU/mL, 38/55, 69% samples with a CV > 1000 IU/mL were detected and 17/55, 30% samples between 12-1000 were not detected. No negative samples (6/6, 100%) were detected by our new qPCR. This in-house real-time PCR showed sensitivity and specificity of 75% and 100% respectively.
This work allowed the local development of a sensitive and efficient qPCR protocol for the detection of samples with CVs elevated (>1000 UI/mL). It will detect pregnant women who need to receive antiviral treatment in order to reduce the risk of HBV transmission. This tool could be extended to other high-risk populations such as immunocompromised people.
乙肝病毒（HBV）的母婴传播（MTCT）是西非等流行地区慢性乙型肝炎的主要病因之一。其预防是消除HBV的重要环节。若无干预措施，HBV垂直传播率因病毒复制水平而异。这种感染的管理是一个主要问题，特别是在像马里这样资源有限的国家，以可承受的成本获取病毒载量。本研究旨在开发并验证一种使用qPCR检测和定量孕妇（有将病毒传播给新生儿风险的人群）中HBV DNA的方法。
我们在本研究中纳入了74例乙肝表面抗原（AgHBs）阳性的孕妇。她们的病毒载量先前已在参考中心实验室测定。我们设计了针对HBV前C基因的特异性探针和引物，用于通过qPCR进行检测和定量。我们对这种新的qPCR进行了调整，以在不同的实时PCR机器上定量HBV DNA。
病毒载量（VL）在10000 - 100000 IU/mL之间的9个样本（9/9，100%）以及VL > 100000 IU/mL的4个样本（4/4，100%）均被检测和定量。在病毒载量为12 - 10000 IU/mL的55个样本中，病毒载量> 1000 IU/mL的38个样本（38/55，69%）被检测到，而12 - 1000之间的17个样本（17/55，30%）未被检测到。我们的新qPCR未检测到任何阴性样本（6/6，100%）。这种内部实时PCR的灵敏度和特异性分别为75%和100%。
这项工作使得能够在当地开发出一种灵敏且高效的qPCR方案，用于检测病毒载量升高（> 1000 UI/mL）的样本。它将检测出需要接受抗病毒治疗以降低HBV传播风险的孕妇。该工具可扩展到其他高危人群，如免疫功能低下者。
