Department of Chemistry, Vanderbilt University, Nashville, Tennessee.
Rusangu University, Monze, Zambia.
Am J Trop Med Hyg. 2018 May;98(5):1389-1396. doi: 10.4269/ajtmh.17-0996. Epub 2018 Mar 15.
A rapid, on-bead enzyme-linked immunosorbent assay for lactate dehydrogenase (LDH) and histidine-rich protein 2 (HRP2) was adapted for use with dried blood spot (DBS) samples. This assay detected both biomarkers from a single DBS sample with only 45 minutes of total incubation time and detection limits of 600 ± 500 pM (LDH) and 69 ± 30 pM (HRP2), corresponding to 150 and 24 parasites/μL, respectively. This sensitive and reproducible on-bead detection method was used to quantify LDH and HRP2 in patient DBS samples from rural Zambia collected at multiple time points after treatment. Biomarker clearance patterns relative to parasite clearance were determined; LDH clearance followed closely with parasite clearance, whereas most patients maintained detectable levels of HRP2 for 35-52 days after treatment. Furthermore, weak-to-moderate correlations between biomarker concentration and parasite densities were found for both biomarkers. This work demonstrates the utility of the developed assay for epidemiological study and surveillance of malaria.
一种快速的、基于珠子的酶联免疫吸附测定法（ELISA）被用于乳酸脱氢酶（LDH）和富含组氨酸蛋白 2（HRP2）的检测，该方法可用于干血斑（DBS）样本。该检测法仅需 45 分钟的总孵育时间，即可从单个 DBS 样本中同时检测到这两种生物标志物，检测限分别为 600±500 pM（LDH）和 69±30 pM（HRP2），相当于 150 和 24 个寄生虫/μL。这种灵敏且可重复的基于珠子的检测方法，用于定量分析赞比亚农村地区患者治疗后多个时间点采集的 DBS 样本中的 LDH 和 HRP2。确定了生物标志物清除与寄生虫清除的相对模式；LDH 的清除与寄生虫的清除密切相关，而大多数患者在治疗后 35-52 天仍可检测到 HRP2。此外，对于两种生物标志物，均发现了生物标志物浓度与寄生虫密度之间的弱到中度相关性。这项工作证明了所开发的检测方法在疟疾的流行病学研究和监测中的实用性。
