Tropical Infectious Diseases Research and Education Centre (TIDREC), University of Malaya, 50603, Kuala Lumpur, Malaysia.
WHO Collaborating Centre for Arbovirus Research and Reference (Dengue and Severe Dengue), University of Malaya, 50603, Kuala Lumpur, Malaysia.
BMC Infect Dis. 2018 Apr 11;18(1):169. doi: 10.1186/s12879-018-3065-1.
A method for rapid detection of dengue virus using the reverse-transcription recombinase polymerase amplification (RT-RPA) was recently developed, evaluated and made ready for deployment. However, reliance solely on the evaluation performed by experienced researchers in a well-structured and well-equipped reference laboratory may overlook the potential intrinsic problems that may arise during deployment of the assay into new application sites, especially for users unfamiliar with the test. Appropriate assessment of this newly developed assay by users who are unfamiliar with the assay is, therefore, vital.
An operational utility test to elucidate the efficiency and effectiveness of the dengue RT-RPA assay was conducted among a group of researchers new to the assay. Nineteen volunteer researchers with different research experience were recruited. The participants performed the RT-RPA assay and interpreted the test results according to the protocol provided. Deviation from the protocol was identified and tabulated by trained facilitators. Post-test questionnaires were conducted to determine the user satisfaction and acceptability of the dengue RT-RPA assay.
All the participants completed the test and successfully interpreted the results according to the provided instructions, regardless of their research experience. Of the 19 participants, three (15.8%) performed the assay with no deviations and 16 (84.2%) performed the assay with only 1 to 5 deviations. The number of deviations from protocol, however, was not correlated with the user laboratory experience. The accuracy of the results was also not affected by user laboratory experience. The concordance of the assay results against that of the expected was at 89.3%. The user satisfaction towards the RT-RPA protocol and interpretation of results was 90% and 100%, respectively.
The dengue RT-RPA assay can be successfully performed by simply following the provided written instructions. Deviations from the written protocols did not adversely affect the outcome of the assay. These suggest that the RT-RPA assay is indeed a simple, robust and efficient laboratory method for detection of dengue virus. Furthermore, high new user acceptance of the RT-RPA assay suggests that this assay could be successfully deployed into new laboratories where RT-RPA was not previously performed.
最近开发了一种使用逆转录重组酶聚合酶扩增（RT-RPA）快速检测登革热病毒的方法，该方法已经过评估并准备部署。然而，仅仅依靠经验丰富的研究人员在结构良好且设备齐全的参考实验室中进行的评估，可能会忽略在将该检测方法应用于新的应用地点时可能出现的潜在内在问题，特别是对于不熟悉该检测方法的用户。因此，不熟悉该检测方法的新用户对该新开发的检测方法进行适当评估至关重要。
在一组不熟悉该检测方法的研究人员中进行了一项操作实用性测试，以阐明登革热 RT-RPA 检测方法的效率和有效性。招募了 19 名具有不同研究经验的志愿者研究人员。参与者根据提供的方案进行 RT-RPA 检测，并根据方案解释检测结果。受过培训的协调员会识别并记录偏离方案的情况。检测后进行问卷调查，以确定用户对登革热 RT-RPA 检测方法的满意度和可接受性。
所有参与者都完成了检测，并根据提供的说明成功解释了结果，无论他们的研究经验如何。在 19 名参与者中，有 3 名（15.8%）在没有任何偏差的情况下进行了检测，而 16 名（84.2%）仅进行了 1 到 5 次偏差的检测。从方案偏离的次数与用户实验室经验无关。用户实验室经验也没有影响检测结果的准确性。检测结果与预期结果的一致性为 89.3%。用户对 RT-RPA 方案和结果解释的满意度分别为 90%和 100%。
只需按照提供的书面说明，即可成功进行登革热 RT-RPA 检测。对书面方案的偏离并未对检测结果产生不利影响。这表明 RT-RPA 检测方法确实是一种简单、稳健且高效的检测登革热病毒的实验室方法。此外，新用户对 RT-RPA 检测方法的高度接受表明，该检测方法可以成功部署到以前未进行 RT-RPA 检测的新实验室。
