Performance Evaluation of RealDetect™ COVID-19 RT-PCR Kit for the Detection of SARS-CoV-2 Virus

NCT04403672 | Unknown

• 1 other identifier: 30005052020
• Study Type: observational
• Target: 120
• Locations: 1 country
• Sites: 1

Brief Summary

The novel Severe acute respiratory syndrome coronavirus 2 (SARS-C0V 2) originated in Wuhan, China in December 2019. As of April 15 2020, the virus has spread across 213 countries/territories with 1,914,916 cases and 123,010 deaths and a crude case fatality ratio (CFR) of 6.4%. In Bangladesh, the situation is also grave. As of May 14, 2020, there were 18,863 cases and 283 deaths. In order to suppress COVID-19 transmission, it is important to diagnose COVID-19 patients, which would help in the process of quarantine and isolation of the patients and also in contact tracing. COVID-19 testing can identify the SARS-CoV-2 virus and includes methods that detect the presence of virus itself such as real time reverse-transcription-polymerase chain reaction (RT-PCR), isothermal nucleic acid amplification, antigen) and those that detect antibodies produced in response to infection. Until now, RT-PCR has been known as the best approach for - detection. It would be very useful if Bangladesh had its own locally produced RT-PCR kits, provided that the kits are no less in quality than imported kits in terms of sensitivity, specificity, price etc. The present study aims to carry out the performance evaluation of RealDetect RT-PCR Kit for COVID-19 detection. The RealDetect™ COVID-19 RT-PCR diagnostic panel is a locally produced real-time RT-PCR test intended for the qualitative detection of nucleic acid from the SARS-CoV-2 in nasopharyngeal swab specimens collected from individuals who meet SARS-CoV-2 clinical criteria. The approach is based on the RT-PCR method which uses two (Nucleocapsid 1, Nucleocapsid 2) sets of gene-specific primers and corresponding fluorescent probes to detect two specific regions within the novel coronavirus (SARS-CoV-2) nucleocapsid protein Nucleocapsid gene. This RT-PCR panel detects SARS-CoV-2 Ribonucleic acid (RNA) specifically. The approach does not generate any false positives to other coronaviruses or human microflora. The kit also contains a primer-probe set which detects human housekeeping gene, ribonuclease Protein (RNase P). That is, the Ribonuclease Protein (RNase P) serves as an internal reference control to monitor sample collection, ribonucleic acid (RNA) extraction, and amplification. This is a case control study. The study 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). These specimens will be blinded before handing over to Institute for Developing Science \& Health Initiatives (ideSHi) for RealDetect Kit. All samples will be analyzed by Real Time PCR System. Necessary validation will also be carried out at the COVID-19 laboratory of the Dhaka Medical College Hospital and an external validation expert will be involved. The Principal Investigator (PI) will also receive the sample information regarding positive/negative status from Institute for Epidemiology, Disease Control and Research (IEDCR) and compare ideSHi and IEDCR data. Unpaired t-test, Wilcox's test, Rank test, Compare test, Mean test, Sensitivity/Specificity test, Regression analysis and Geometric mean with 95% Confidence Interval (CI) will be used to analyze the data. It needs to conduct a univariate analysis.

Conditions

High Sensitivity and Specificity (With 95% Confidence Interval) of RealDetect™ COVID-19 RT-PCR Kit

Outcome Measures

Primary Outcomes (1)

• Performance evaluation of RealDetect™ COVID-19 RT-PCR kit

  Determine Performance evaluation of RealDetect™ COVID-19 RT-PCR kit for the detection of SARS-CoV-2 virus using nasopharyngeal swab specimens collected in the nationwide COVID-19 screening program. RNA extraction from fresh Nasopharyngeal Swab sample in Viral Transport Medium(VTM) of COVID-19 patients from IEDCR (30 positive \& 30 negative) Analysis of COVID-19 RNA samples using RT-PCR

  2 months

Secondary Outcomes (1)

• Reduce the price of RT-PCR based COVID-19 diagnostic test kits.

  1 month

Study Arms (2)

COVID-19 positive

ideSHi (CRO) investigators will be blinded to the samples provided by IEDCR. That is, ideSHi investigators will not know which of the 60 samples are positive and which of the 60 samples are negative. After RT- PCR run using RealDetect RT-PCR Kit, ideSHi will send the results to the Principal investigator. The Principal investigator will also receive IEDCR results for the same samples. Finally, the PI will compare the results from ideSHi and IEDCR and determine the sensitivity, specificity, positive predictive value and negative predictive value.

Device: performance evaluation study of RealDetect RT-PCR Kit for COVID-19 detection

COVID-19 Negative

ideSHi investigators will be blinded to the samples provided by IEDCR. That is, ideSHi investigators will not know which of the 60 samples are positive and which of the 60 samples are negative. After RT- PCR run using RealDetect RT-PCR Kit, ideSHi will send the results to the Principal investigator. The Principal investigator will also receive IEDCR results for the same samples. Finally, the PI will compare the results from ideSHi and IEDCR and determine the sensitivity, specificity, positive predictive value and negative predictive value. In addition, 60 fresh specimens which will be tested at IEDCR will also be tested at ideSHi using RealDetect on a real time basis. These samples will also be blinded by IEDCR and sent to the testing laboratory (ideSHi). Of these 60 samples, 30 will be COVID-19 positive and 30 COVID- 19 negative samples. These fresh samples will be provided to ideSHi for testing and analysis for performance evaluation of RealDetect COVID-19 RT-PCR kit.

Device: performance evaluation study of RealDetect RT-PCR Kit for COVID-19 detection

Interventions

performance evaluation study of RealDetect RT-PCR Kit for COVID-19 detection DEVICE

IEDCR uses many different kits for COVID-19 detection. These kits include, (1) A\*Star Fortitute Kit 2 for COVID-19 detection (Singapure), (2) Sansure Biotec Limited (China), (3) ModularDx Kit (TIB Molecular Biology, Germany, and (4) DaAn Kit (China). Among these 2 kits, A\*Star Fortitude Kit 2 has been known as the best kit in IEDCR depending on its performance because it rarely generates false positive or false negative results. A\*Star Fortitude Kit 2 is used for confirmation of any discordant results for other kits in IEDCR. That is, IEDCR uses A\*Star Fortitude Kit for COVID detection as the Gold Standard. In our case, this is the main reason for the selection of this kit in order to compare the performances of RealDetect™ COVID-19 RT-PCR kit.

COVID-19 Negative; COVID-19 positive

Eligibility Criteria

• Age: 5 Years - 65 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17), Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

Testing and analysis of a total of 120 specimens will be carried out. Of the 120 samples, 60 will be stored samples (30 COVID-19 positive samples and 30 COVID-19 negative samples) from IEDCR. The positive/negative status of the samples will be blinded before sending to ideSHi. The samples have already been stored at -80 degree Celsius in the IEDCR freezer. In addition, 60 fresh specimens which will be tested at IEDCR will also be tested at ideSHi using RealDetect on a real time basis. These samples will also be blinded by IEDCR and sent to the testing laboratory (ideSHi) as they become available. Of these 60 samples, 30 will be COVID-19 positive and 30 COVID- 19 negative samples. These fresh samples will be provided to ideSHi for testing and analysis for performance evaluation of RealDetect COVID-19 RT-PCR kit. The PI will coordinate these activities.

You may qualify if:

• Samples with positive RT-PCR results with Ct value ≤30 for the COVID-19 gene(s) at IEDCR will be selected as COVID-19 positive.
• Samples with negative RT-PCR results with no amplification for the COVID-19 gene(s) at IEDCR will be selected as COVID-19 negative.

You may not qualify if:

• Samples with equivocal/ambiguous RT-PCR results in terms of sigmoidal curve and Ct value will be excluded.

Sponsors & Collaborators

• Bangladesh Medical Research Council (BMRC): lead

Study Sites (1)

Institute of Epidemiology, Disease Control and Research (IEDCR), Mohakhali, Dhaka (For sample collection & sample storage), Institute for Developing Science & Health Initiatives (ideSHi); Mohakhali, Dhaka (For sample analysis, data collection, data analys

Dhaka, Bangladesh

RECRUITING

Related Publications (2)

• Coronaviridae Study Group of the International Committee on Taxonomy of Viruses. The species Severe acute respiratory syndrome-related coronavirus: classifying 2019-nCoV and naming it SARS-CoV-2. Nat Microbiol. 2020 Apr;5(4):536-544. doi: 10.1038/s41564-020-0695-z. Epub 2020 Mar 2.

  PMID: 32123347 BACKGROUND

• Hui DS, I Azhar E, Madani TA, Ntoumi F, Kock R, Dar O, Ippolito G, Mchugh TD, Memish ZA, Drosten C, Zumla A, Petersen E. The continuing 2019-nCoV epidemic threat of novel coronaviruses to global health - The latest 2019 novel coronavirus outbreak in Wuhan, China. Int J Infect Dis. 2020 Feb;91:264-266. doi: 10.1016/j.ijid.2020.01.009. Epub 2020 Jan 14. No abstract available.

  PMID: 31953166 BACKGROUND

Related Links

• Novel Coronavirus (2019-nCoV): Situation Report
• Coronavirus disease 2019 (COVID-19) Situation Report-85

Biospecimen

Retention: SAMPLES WITH DNA

Nasopharyngeal swabs

Study Officials

• Zahid Hassan, MBBS, PhD

  Bangladesh University of Health Sciences

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Rosy Sultana, MBBS, MPhil

CONTACT

+88 01711014806; sultanarosy64@yahoo.com, sultanarosy@buhs.ac.bd

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: PROSPECTIVE
• Target Duration: 2 Months
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: May 24, 2020
• First Posted: May 27, 2020
• Study Start: May 18, 2020
• Primary Completion: July 24, 2020
• Study Completion: July 24, 2020
• Last Updated: May 28, 2020

Record last verified: 2020-05

Data Sharing

• IPD Sharing: Will not share

Locations

BA (1)