Development of a Multiplex BangasureTM RT-PCR Kit for the Detection of SARS-Corona Virus-2
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 150
- 试验地点
- 1
- 主要终点
- Performance evaluation of BRiCM COVID-19 RT-PCR
研究概览
简要总结
The outbreak of the severe acute respiratory syndrome (SARS) corona virus (CoV)-2 continues to increase globally. To control the coronavirus disease 2019 (COVID-19) transmission, diagnose is very important step which would help in the process of quarantine, isolation of the infected person and also in contact tracing. Among the various techniques real-time reverse transcription polymerase chain reaction (rRT-PCR) is mostly used and considers as gold standard method for viral nucleic acid detection as per World Health Organization (WHO) and Centre for Diseases Control (CDC), USA. However, possible false-negative and false-positive results produce misleading consequences, making it necessary to improve existing methods. So, investigators need a robust and reliable PCR kit to detect different kinds of variant of SARS-CoV-2 virus to reduce its transmissibility and take preventive management plan. Beside that to manage the increasing number of testing, investigators are mostly dependent on imported test kit which seems to be very difficult at this pandemic situation and costly. Hence, virulence of SARS-CoV-2 virus may change rapidly due to mutation and country become hot spot so, gene detection RT-PCR kit is time demand. In this point of view, investigator's aim to develop an validate multiplex rRT-PCR qualitative diagnostic method, which targets two viral genes E (envelope protein), N (nucleocapsid gene, according to the CDC, USA guidelines for the development of RT-PCR kit and one human gene RNase P as internal control simultaneously.
This is a case control study where will analyze 120 samples (60 COVID-19 positive and 60 COVID-19 negative both fresh and frozen) from Institute of Epidemiology, Disease Control and Research (IEDCR). This specimen will be blinded before supplying over to NILMRC/Contact ROL/ Bangladesh Clinical Trail Limited (BCTL), Dhaka, Bangladesh for Bangladesh Reference Institute for Chemical Measurements (BRiCM) RT-PCR Kit. Required in house validation will also be conducted as per International Council for Harmonization (ICH) and FDA guideline and also external validation will be carried in different Director General of Health Service (DGHS), Bangladesh approved COVID-19 diagnostic laboratory. Data will be analyzed by computer using Statistical Package for the Social Sciences (SPSS) software and will be presented into the frequency table, graph and chart according to the requirements.
详细描述
Introduction:
The corona virus COVID-19 pandemic is the defining global health crisis of our time and the greatest challenge that have faced since World War Two. The pandemic is much more than a health crisis; it's also an unprecedented socio-economic crisis. Since its identification in the region of Wuhan, China, not only the virus has spread across 213 countries/territories with 209 million cases and 4.39 million globally but also could put 400million people out of their jobs as The International Labor Organization estimates.
To contain the spread of disease, multidisciplinary strategies have been launched in different regions of the world, including implementing social distancing, maintaining personal hygiene, contact tracing, and implementing quarantine, travel restrictions, and lock downs. A widely accepted method is testing for SARS-CoV-2, typically utilizing nasopharyngeal swab specimens. Patients tested positive require appropriate clinical management by either effective isolation or quarantine at home for mild symptoms or within health care facilities for moderate to severe symptoms. In addition, wide-scale testing provides more informative epidemiologic data for drafting policies on disease monitoring and control. Many approaches have been proposed to detect SARS-CoV-2 virus in nasopharyngeal fluids such as multiplex RT-PCR, CRISPR/Cas12, and CRISPR-Cas3, lateral flow immunoassay, paper-based bimolecular sensors, SHERLOCK testing in one pot, DNA aptamer, loop-mediated isothermal amplification (LAMP), etc. Among these methods, nucleic acid amplification-based tests are the most common for the diagnosis of SARS-CoV-2.
In addition, US CDC suggest a protocol for the detection of SARS-CoV-2, based on the amplification of two regions of the nucleocapsid gene, namely N1and N2, and a human internal control gene RNase P (RP). In addition to these strategies, efforts to develop SARS-CoV-2 detection methods with high efficiency and accuracy, with less reaction time and effort, are still ongoing.
However, despite being considered as the gold standard for SARS CoV-2 detection, testing has lagged behind in many countries because of various factors, most important being supply chain issues with lack of reagents and adequate test kits. Therefore, many patients (both symptomatic and asymptomatic) remain untested and hence are potentially contributing to community spread of the virus.
研究设计
- 研究类型
- Observational
- 观察模型
- Case Control
- 时间视角
- Prospective
入排标准
- 年龄范围
- 5 Years 至 70 Years（Child, Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •Samples with Positives RT-PCR results with Ct value≤35 for the COVID-19 genes at IEDCR will be selected as COVID-19 positive.
- •Samples with negative RT-PCR results with no amplification for the COVID-19 genes at IEDCR will be selected as COVID-19.
排除标准
- •Samples with equivocal/ambiguous RT-PCR results in terms of sigmoidal curve and Ct value will be excluded.
结局指标
主要结局
Performance evaluation of BRiCM COVID-19 RT-PCR
时间窗: 1 month
Determine Performance evaluation of BRiCM COVID-19 RT-PCR with extracted viral RNA by using standard viral RNA extraction kit for the detection of SARS-CoV-2 virus in nasopharyngeal swab specimens collected in the nationwide COVID-19 screening program. RNA extraction from fresh Nasopharyngeal Swab sample in the Viral Transport Medium (VTM) of COVID-19 patients from IEDCR (30 positive and 30 negative) Analysis of COVID-19 RNA samples using RT-PCR.
次要结局
- Reduce the price and enhance the efficiency to detect COVID-19 by BRiCM COVID-19 RT-PCR kit(1 month)
研究者
DR. MALA KHAN
Director General
Bangladesh Reference Institute of Chemical Measurements (BRICM)