Infectious and Tropical Diseases Research Group (e-INTRO), Faculty of Pharmacy, Biomedical Research Institute of Salamanca-Research Centre for Tropical Diseases at the University of Salamanca (IBSAL-CIETUS), University of Salamanca, Salamanca, Spain.
Department of Parasitology, National Centre for Microbiology, Institute of Health Carlos III, Majadahonda, Madrid, Spain.
Trop Med Int Health. 2018 Sep;23(9):992-1001. doi: 10.1111/tmi.13117. Epub 2018 Jul 5.
To evaluate the performance of Rapid-Heat LAMPellet assay in field conditions for diagnosis of urogenital schistosomiasis in an endemic area in Cubal, Angola, and to assess the reproducibility in a reference laboratory.
A total of 172 urine samples from school-age children were tested for microhaematuria, microscopic detection of Schistosoma haematobium eggs and LAMP for DNA detection. Urine samples were stored in a basic equipped laboratory. Field-LAMP tests were performed with and without prior DNA extraction from urine samples, and the results were read by turbidity and by colour change. When field procedures were finished, samples were sent to a reference laboratory to be reanalysed by LAMP.
A total of 83 of 172 (48.3%) were positive for microhaematuria, 87/172 (50.6%) were microscopy-positive for S. haematobium eggs detection, and 127/172 (73.8%) showed LAMP-positive results for detecting S. haematobium using purified DNA and 109/172 (63.4%) without prior DNA extraction. MacNemar's test showed a statistical significant relation between LAMP results and microscopy-detected S. haematobium infections and microhaematuria (P < 0.001 in both cases), respectively. When samples of purified DNA were reanalysed in a reference laboratory in Spain using the same LAMP methodology, the overall reproducibility achieved 72.1%.
The ease of use, simplicity and feasibility demonstrated by LAMP assay in field conditions together with the acceptable level of reproducibility achieved in a reference laboratory support the use of LAMP assay as an effective test for molecular diagnosis of urogenital schistosomiasis in endemic remote areas.
评估 Rapid-Heat LAMPellet assay 在安哥拉 Cubal 流行地区现场条件下用于诊断泌尿生殖系统血吸虫病的性能，并评估在参考实验室中的重现性。
共检测了 172 份来自学龄儿童的尿液样本，以检测镜下血尿、镜下检测 Schistosoma haematobium 卵和 DNA 检测的 LAMP。尿液样本储存在一个基本配备的实验室中。在现场进行 LAMP 测试时，尿液样本分别进行了有和无 DNA 提取的检测，并通过浊度和颜色变化来读取结果。当现场程序完成后，将样本送到参考实验室，用 LAMP 重新分析。
172 份样本中，共有 83 份（48.3%）对镜下血尿呈阳性，87 份（50.6%）对 S. haematobium 卵检测呈显微镜阳性，127 份（73.8%）用纯化 DNA 检测 S. haematobium 的 LAMP 结果呈阳性，109 份（63.4%）未进行 DNA 提取。MacNemar 检验显示，LAMP 结果与显微镜检测到的 S. haematobium 感染和镜下血尿之间存在统计学显著关系（两种情况的 P 值均<0.001）。当在西班牙的参考实验室用相同的 LAMP 方法对纯化的 DNA 样本进行重新分析时，总体重现性达到 72.1%。
LAMP assay 在现场条件下的易用性、简单性和可行性，以及在参考实验室中达到的可接受的重现性水平，支持将 LAMP assay 作为一种有效的分子诊断泌尿生殖系统血吸虫病的方法，用于在偏远的流行地区。
