Kituyi Sarah N, Nyakoe Nancy, Ngeranwa Joseph N, Runo Steven, Waitumbi John N
Biochemistry and Biotechnology Department, Kenyatta University, Nairobi, Kenya.
Walter Reed Project, Kenya Medical Research Institute, Kisumu, Kenya.
Malariaworld J. 2014 Jun 26;5:7. doi: 10.5281/zenodo.10887755. eCollection 2014.
Accurate diagnosis of malaria is key to proper management and control and an ideal diagnostic parameter that correlates to disease outcome is required. The former would be helpful in correctly identifying patients that need hospitalisation versus those that can be managed at home. This study determined how well the density estimates by microscopy, qPCR and HRP-2 correlate to malaria severity.
Patients aged ≤ 5 yrs with severe (n = 60, Hb ≤ 6 g/dl) and mild (n = 60, Hb > 6 g/dl) malaria were enrolled to take part in a case control study at Kisumu District Hospital, Western Kenya. Parasite load was determined by microscopy, qPCR targeting the rRNA gene and HRP-2 antigen ELISA.
The median parasite load and the 25 and the 75 percentile by microscopy in children with severe malaria (SM) was 49,958 parasites/μl (12,013-128,695) compared to 24,233 (6,122-103,886) in the group with mild malaria (MM), P = 0.10. By qPCR, the translated median parasite density was 31,550 parasites/μl (4,106-196,640) in the SM group compared to 24,365 parasites/μl (5,512-93,401) in the MM group (P = 0.73). According to HRP-2, the translated median parasite load in children with SM was 628,775 parasites/μl (332,222-1.165x106) compared to 150,453 (94,292-399,100) in children with MM (P < 0.0001).
Unlike microscopy and qPCR, the parasite load detected by HRP-2 correlates with disease severity. Because of its unique attributes, HRP-2 is able to account for trophozoites and schizonts that are sequestered away from peripheral circulation. Because it persists in circulation, it also serves as an indicator of the magnitude of current and recent infections.
疟疾的准确诊断是合理管理与控制的关键,需要一个与疾病结局相关的理想诊断参数。前者有助于正确识别需要住院治疗的患者与可在家中治疗的患者。本研究确定了显微镜检查、定量聚合酶链反应(qPCR)和疟原虫富含组氨酸蛋白2(HRP-2)检测的密度估计值与疟疾严重程度的相关性。
年龄≤5岁的重度(n = 60,血红蛋白≤6 g/dl)和轻度(n = 60,血红蛋白>6 g/dl)疟疾患者在肯尼亚西部基苏木区医院参加了一项病例对照研究。通过显微镜检查、针对核糖体RNA(rRNA)基因的qPCR和HRP-2抗原酶联免疫吸附测定(ELISA)确定寄生虫载量。
重度疟疾(SM)患儿通过显微镜检查测得的寄生虫载量中位数以及第25和第75百分位数为49,958个寄生虫/微升(12,013 - 128,695),而轻度疟疾(MM)组为24,233个(6,122 - 103,886),P = 0.10。通过qPCR,SM组的寄生虫密度中位数经换算为31,550个寄生虫/微升(4,106 - 196,640),MM组为24,365个寄生虫/微升(5,512 - 93,401)(P = 0.73)。根据HRP-2检测结果,SM患儿的寄生虫载量中位数经换算为628,775个寄生虫/微升(332,222 - 1.165×10⁶),MM患儿为150,453个(94,292 - 399,100)(P < 0.0001)。
与显微镜检查和qPCR不同,HRP-2检测到的寄生虫载量与疾病严重程度相关。由于其独特的特性,HRP-2能够检测到隔离于外周循环之外的滋养体和裂殖体。由于它在循环中持续存在,它还可作为当前和近期感染程度的指标。