ICMR-National Institute of Malaria Research, New Delhi, India.
Kumaun University, Nainital, Uttarakhand, India.
J Vector Borne Dis. 2023 Apr-Jun;60(2):200-206. doi: 10.4103/0972-9062.374038.
BACKGROUND & OBJECTIVES: The highly sensitive method for a true understanding of malaria prevalence is of utmost importance for India's elimination strategy. The PCR reaction type with rapid detection, cost-effectiveness, and less workforce should be preferable. Multiplex PCR type accomplishes the present requirement by saving time and resources to find true surveillance data for malaria, especially in low-parasitemia/asymptomatic groups or populations.
The present study focuses on designing multiplex PCR (mPCR) to detect simultaneously Plasmodium genus (PAN) and two common Plasmodium species found in India. It is compared to standard nested PCR on 195 clinical samples to diagnose malaria. The mPCR was designed with a minimum number of primers, leading to less clogging and effective and enhanced detection. It contains one common reverse primer and three forward primers amplifying three targeted genes corresponding to P. falciparum, P. vivax, and Plasmodium genus.
The sensitivity and specificity for mPCR were 94.06 and 95.74, respectively. The limit of detection for mPCR was 0.1 parasites/µl. The study has shown a ROC curve area for the mPCR of 0.949 for Plasmodium genus and P. falciparum and 0.897 for P. vivax with standard nPCR.
INTERPRETATION & CONCLUSION: The mPCR is rapid in detecting species together, cost-effective, and requires fewer human resources than the standard nPCR. Therefore, the mPCR can be used as an alternative technique for the higher sensitive detection of the malaria parasite. It could also become a vital tool for determining malaria prevalence, facilitating the application of the most effective measures.
对于印度的消除战略而言，采用高度敏感的方法来真正了解疟疾的流行情况至关重要。PCR 反应类型具有快速检测、具有成本效益且所需劳动力较少，因此更可取。多重 PCR 类型通过节省时间和资源来寻找疟疾的真实监测数据，特别是在低寄生虫血症/无症状组或人群中，从而满足了当前的要求。
本研究旨在设计多重 PCR（mPCR）来同时检测在印度发现的疟原虫属（PAN）和两种常见的疟原虫。它与标准巢式 PCR 一起用于诊断 195 例临床样本中的疟疾。mPCR 的设计使用了尽可能少的引物，从而减少了堵塞并提高了检测的有效性和增强了检测效果。它包含一个通用的反向引物和三个正向引物，可扩增三个靶向基因，分别对应于恶性疟原虫、间日疟原虫和疟原虫属。
mPCR 的灵敏度和特异性分别为 94.06%和 95.74%。mPCR 的检测限为 0.1 个寄生虫/μl。该研究表明，mPCR 对疟原虫属和恶性疟原虫的 ROC 曲线面积为 0.949，对间日疟原虫的 ROC 曲线面积为 0.897，而标准 nPCR 的 ROC 曲线面积分别为 0.923 和 0.863。
mPCR 能够快速同时检测多种物种，具有成本效益，并且所需的人力资源比标准 nPCR 少。因此，mPCR 可以作为一种替代技术用于更敏感地检测疟原虫。它也可以成为确定疟疾流行情况的重要工具，从而有助于实施最有效的措施。
