Abstract：
UNASSIGNED: Plasmodium knowlesi, a simian malaria species, is now known to infect humans. Due to disadvantages in the current diagnosis methods, many efforts have been placed into developing new methods to diagnose the disease. This study assessed the ability of the PkRAP-1 sandwich enzyme-linked immunosorbent (ELISA) to detect P knowlesi antigens in whole blood specimens.
METHODS: Western blot assay was conducted to evaluate the ability of raised mouse and rabbit anti-PkRAP-1 polyclonal antibodies to bind to the native proteins in P. knowlesi lysate. The polyclonal antibodies were then used in sandwich ELISA to detect P. knowlesi. In the sandwich ELISA, mouse and rabbit polyclonal antibodies were used as the capture and detection antibodies, respectively. The limit of detection (LOD) of the assay was determined using P. knowlesi A1H1 culture and purified recombinant PkRAP-1.
RESULTS: Western blot results showed positive reactions towards the proteins in P. knowlesi lysate. The LOD of the assay from three technical replicates was 0.068% parasitaemia. The assay performance in detecting P. knowlesi was 83% sensitivity and 70% specificity with positive and negative predictive values of 74% and 80%, respectively. The anti-PkRAP-1 polyclonal antibodies did not cross-react with P. falciparum and healthy samples, but P. vivax by detecting all 12 samples.
UNASSIGNED: PkRAP-1 has the potential as a biomarker for the development of a new diagnostic tool for P. knowlesi detection. Further studies need to be conducted to establish the full potential of the usage of anti-PkRAP-1 antibodies for P. knowlesi detection.
METHODS: Western blot assay was conducted to evaluate the ability of raised mouse and rabbit anti-PkRAP-1 polyclonal antibodies to bind to the native proteins in P. knowlesi lysate. The polyclonal antibodies were then used in sandwich ELISA to detect P. knowlesi. In the sandwich ELISA, mouse and rabbit polyclonal antibodies were used as the capture and detection antibodies, respectively. The limit of detection (LOD) of the assay was determined using P. knowlesi A1H1 culture and purified recombinant PkRAP-1.
RESULTS: Western blot results showed positive reactions towards the proteins in P. knowlesi lysate. The LOD of the assay from three technical replicates was 0.068% parasitaemia. The assay performance in detecting P. knowlesi was 83% sensitivity and 70% specificity with positive and negative predictive values of 74% and 80%, respectively. The anti-PkRAP-1 polyclonal antibodies did not cross-react with P. falciparum and healthy samples, but P. vivax by detecting all 12 samples.
UNASSIGNED: PkRAP-1 has the potential as a biomarker for the development of a new diagnostic tool for P. knowlesi detection. Further studies need to be conducted to establish the full potential of the usage of anti-PkRAP-1 antibodies for P. knowlesi detection.
摘要：
■诺氏疟原虫，一种类人猿疟疾，现在已知会感染人类。由于当前诊断方法的缺点，许多努力已经投入到开发新的方法来诊断这种疾病。这项研究评估了PkRAP-1夹心酶联免疫吸附（ELISA）检测全血标本中Pknowlesi抗原的能力。
方法:进行Western印迹测定以评估提高的小鼠和兔的抗PkRAP-1多克隆抗体结合诺氏磷裂解物中的天然蛋白的能力。然后在夹心ELISA中使用多克隆抗体来检测诺氏假单胞菌。在夹心ELISA中,小鼠和兔多克隆抗体被用作捕获和检测抗体，分别。该测定的检测限(LOD)是使用诺氏假单胞菌A1H1培养物和纯化的重组PkRAP-1测定的。
结果：Western印迹结果显示，对诺氏磷裂解物中的蛋白质呈阳性反应。来自三个技术重复的测定的LOD为0.068%寄生虫血症。检测诺氏假单胞菌的测定性能为83%的灵敏度和70%的特异性，阳性和阴性预测值分别为74%和80%。分别。抗PkRAP-1多克隆抗体不与恶性疟原虫和健康样本发生交叉反应，但是通过检测所有12个样本来检测间日疟原虫。
■PkRAP-1具有作为生物标志物的潜力，可用于开发一种新的诺氏疟原虫检测诊断工具。需要进行进一步的研究，以确定使用抗PkRAP-1抗体进行知识假单胞菌检测的全部潜力。
方法:进行Western印迹测定以评估提高的小鼠和兔的抗PkRAP-1多克隆抗体结合诺氏磷裂解物中的天然蛋白的能力。然后在夹心ELISA中使用多克隆抗体来检测诺氏假单胞菌。在夹心ELISA中,小鼠和兔多克隆抗体被用作捕获和检测抗体，分别。该测定的检测限(LOD)是使用诺氏假单胞菌A1H1培养物和纯化的重组PkRAP-1测定的。
结果：Western印迹结果显示，对诺氏磷裂解物中的蛋白质呈阳性反应。来自三个技术重复的测定的LOD为0.068%寄生虫血症。检测诺氏假单胞菌的测定性能为83%的灵敏度和70%的特异性，阳性和阴性预测值分别为74%和80%。分别。抗PkRAP-1多克隆抗体不与恶性疟原虫和健康样本发生交叉反应，但是通过检测所有12个样本来检测间日疟原虫。
■PkRAP-1具有作为生物标志物的潜力，可用于开发一种新的诺氏疟原虫检测诊断工具。需要进行进一步的研究，以确定使用抗PkRAP-1抗体进行知识假单胞菌检测的全部潜力。
