Department of Public Health, College of Health Sciences, Bahir Dar University, Bahir Dar, Ethiopia.
Department of Microbiology, Immunology and Parasitology, Addis Ababa University, Addis Ababa, Ethiopia.
PLoS One. 2022 Aug 4;17(8):e0272094. doi: 10.1371/journal.pone.0272094. eCollection 2022.
Malaria elimination effort is hampered not only by the lack of effective medication but also due to the lack of sensitive diagnostic tools to detect infections with low levels of parasitemia. Therefore, more sensitive and specific high-throughput molecular diagnostic approaches are needed for accurate malaria diagnosis.
In the present study, the performance of a novel single-tube MC004 real-time polymerase chain reaction (PCR) assay (MRC-Holland, Amsterdam, the Netherlands) was assessed for the detection of infection and discrimination of Plasmodium species. Blood samples (n = 150) were collected from malaria suspected patients at Adama malaria diagnosis and treatment centre, Adama, central Ethiopia. The positive predictive value (PPV), negative predictive value (NPV), analytical sensitivity and specificity of the assay were assessed against the conventional microscopic method.
Plasmodium species were detected in 59 (39.3%) of the samples by microscopy and in 62 (41.3%) by the novel MC004 RT-PCR. Plasmodium vivax, Plasmodium falciparum and mixed infections with Plasmodium falciparum & Plasmodium vivax accounted for 47.5%, 40.6% and 11.9% respectively as detected by microscopy. The MC004 RT-PCR assay identified 59.7% and 40.3% of the samples positive for Plasmodium vivax and Plasmodium falciparum respectively. The sensitivity, specificity, PPV, and NPV of the MC004 RT-PCR assay were 95.8%, 97.8%, 92%, and 98.9%, respectively. No mixed infections were detected using the MC004 assay.
The MC004 RT-PCR assay is a useful tool for the early detection of malaria and identification of Plasmodium species with a high degree of sensitivity and specificity. Due to its high sensitivity, and simplicity (being a single-tube assay), the MC004 is suitable for use in clinical settings and epidemiological studies.
疟疾消除工作不仅受到缺乏有效药物的阻碍，还受到缺乏灵敏诊断工具来检测低疟原虫血症感染的阻碍。因此，需要更灵敏和特异的高通量分子诊断方法来进行准确的疟疾诊断。
本研究评估了一种新型单管 MC004 实时聚合酶链反应（PCR）检测（荷兰马斯特里赫特 MRC 公司）在检测感染和区分疟原虫种方面的性能。从埃塞俄比亚中部阿达玛疟疾诊断和治疗中心的疑似疟疾患者中采集了 150 份血样。该检测方法对常规显微镜法的阳性预测值（PPV）、阴性预测值（NPV）、分析灵敏度和特异性进行了评估。
显微镜法检测到 59 份（39.3%）样本有疟原虫，新型 MC004 RT-PCR 法检测到 62 份（41.3%）样本有疟原虫。显微镜法检测到的疟原虫种分别为间日疟原虫 47.5%、恶性疟原虫 40.6%和恶性疟原虫与间日疟原虫混合感染 11.9%。MC004 RT-PCR 检测到 59.7%和 40.3%的样本为间日疟原虫和恶性疟原虫阳性。MC004 RT-PCR 检测的灵敏度、特异性、PPV 和 NPV 分别为 95.8%、97.8%、92%和 98.9%。MC004 检测未发现混合感染。
MC004 RT-PCR 检测法是一种有用的早期诊断疟疾和鉴定疟原虫种的工具，具有高度的灵敏度和特异性。由于其高灵敏度和简便性（为单管检测），MC004 适用于临床和流行病学研究。
