Abstract：
BACKGROUND: Prompt malarial treatment and surveillance is crucial for accurate diagnosis of Plasmodium Sp. Gold standard microscopic examination has been widely applied for diagnosis of malaria in most part of the endemic areas. But in case of submicroscopic and asymptomatic microscopic diagnosis is questioned. The study aims to develop a simple, cost effective & robust nucleic acid amplification technique for the detection of malaria parasite.
METHODS: Study population included 50 clinically diagnosed positive malaria patient samples from various pathological laboratories. Microscopy by preparing thick film was carried out of every sample for primary screening in the available facility of Surat Raktadan Kendra & Research Centre- Blood Bank. The conventional PCR (Polymerase Chain Reaction) was applied for genus-specific amplification targeting the 18 S rRNA gene of Plasmodium. Agarose gel electrophoresis was used to separate and analyze the amplified PCR product using 2% Agarose gel.
CONCLUSIONS: The study shows that nested PCR not only detected all microscopic positive samples, but also detected submicroscopic infections that were missed or misread by microscopy. Hence, the sensitivity of molecular based detection technique is proved to be more compared to microscopic examination.
METHODS: Study population included 50 clinically diagnosed positive malaria patient samples from various pathological laboratories. Microscopy by preparing thick film was carried out of every sample for primary screening in the available facility of Surat Raktadan Kendra & Research Centre- Blood Bank. The conventional PCR (Polymerase Chain Reaction) was applied for genus-specific amplification targeting the 18 S rRNA gene of Plasmodium. Agarose gel electrophoresis was used to separate and analyze the amplified PCR product using 2% Agarose gel.
CONCLUSIONS: The study shows that nested PCR not only detected all microscopic positive samples, but also detected submicroscopic infections that were missed or misread by microscopy. Hence, the sensitivity of molecular based detection technique is proved to be more compared to microscopic examination.
摘要：
背景：迅速的疟疾治疗和监测对于准确诊断疟原虫Sp至关重要。金标准显微镜检查已广泛应用于大部分流行地区的疟疾诊断。但在亚显微和无症状的情况下，显微镜诊断是有疑问的。该研究旨在开发一种简单的，具有成本效益和强大的核酸扩增技术，用于检测疟原虫。
方法：研究人群包括来自不同病理实验室的50例临床诊断为阳性的疟疾患者样本。在SuratRaktadanKendra&ResearchCenter-血库的现有设施中,通过制备厚膜对用于初级筛选的每个样品进行显微镜检查。将常规PCR(聚合酶链反应)用于靶向疟原虫18SrRNA基因的属特异性扩增。使用琼脂糖凝胶电泳分离和使用2%琼脂糖凝胶分析扩增的PCR产物。
结论：研究表明，巢式PCR不仅检测到所有的显微镜阳性样品，但也检测到显微镜检查遗漏或误读的显微镜下感染。因此，与显微镜检查相比，基于分子的检测技术的灵敏度更高。
方法：研究人群包括来自不同病理实验室的50例临床诊断为阳性的疟疾患者样本。在SuratRaktadanKendra&ResearchCenter-血库的现有设施中,通过制备厚膜对用于初级筛选的每个样品进行显微镜检查。将常规PCR(聚合酶链反应)用于靶向疟原虫18SrRNA基因的属特异性扩增。使用琼脂糖凝胶电泳分离和使用2%琼脂糖凝胶分析扩增的PCR产物。
结论：研究表明，巢式PCR不仅检测到所有的显微镜阳性样品，但也检测到显微镜检查遗漏或误读的显微镜下感染。因此，与显微镜检查相比，基于分子的检测技术的灵敏度更高。
