Regulation/Guidelines

Accelerated Approval Of RealCycler CORO-G Real Time PCR Kits By CliniExperts

  • 2nd September, 2020
Accelerated Approval of RealCycler CORO-G Real Time PCR Kits by CliniExperts

The rapid spread and alarming increase in the number of coronavirus disease 19 (COVID-19) cases caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the community is increasing the burden in economy and healthcare system. The rapid and accurate detection of viruses is increasingly vital to control the sources of infection and help patients prevent further disease progression. Since December 2019, after the outbreak of the SARS-CoV-2in Hubei province of China, there has been a considerable challenge among healthcare professionals regarding the use of nucleic acid tests or clinical characteristics of infected patients as the reference standard make a definitive diagnose of COVID-19 patients. For prevention and control of this pandemic, the early diagnosis of COVID-19 is critical .Patients presenting early-onset of symptoms, clinical characteristics cannot alone define the diagnosis of COVID-19.

Along with advancements in medical diagnosis, nucleic acid detection-based approaches have become a rapid and reliable technology for the detection of viruses. The polymerase chain reaction (PCR) method is considered the ‘gold standard’ for detecting viruses among nucleic acid tests. It is characterized by rapid detection, high sensitivity, and specificity. Progenie molecular from Spain recently got approval from the Central Drugs Standard Control Organisation (CDSCO) for precise results of RealCycler CORO-G Real-Time PCR Kits, helping in the diagnosis of India’s corona cases.

What are RealCycler CORO-G Kits v.5.3?

RealCycler CORO-G Kits v.5.3 is Real-time PCR Test Kit, using Taqman-like hydrolysis probes technology, in combination with CHIC technology for the design of the internal amplification control.

The kits allows the detection of Coronavirus SARS-CoV-2 and Sarbecovirus E gene RNA in clinical samples using realtime PCR

The system includes an internal control of amplification based on the technology CHIC (Competitive Heterologous Internal Control) to prevent false negatives due to reaction inhibition. In Chic-Out RealCycler kits, CHIC is out of the AmpliMix to verify the nucleic acid extraction.

A Positive Control is included in the ReaCycler, which consists of a DNA artificial construction of Coronavirus, is included in the ReaCycler

Where can we use RealCycler CORO-G v.5.3?

RealCycler CORO-G v.5.3 is an in vitro diagnostic kit of reagents which allows the detection of Coronavirus SARS-CoV-2 and Sarbecovirus RNA in clinical samples

  • It is mainly targeted by ORF8 (specific of SARS-CoV-2) + E gene (Sarbecovirus)
  • Samples: It is suitable for the in-vitro qualitative detection of SARS-CoV-2 nucleic acids in oropharyngeal swabs, nasopharyngeal swabs and nasal swabs collected from individuals suspected of COVID 19 by their healthcare provider (HCP). Samples including from individuals without symptoms or other reasons to suspect COVID-19 infection.
  • RealCycler CORO-G is compatible with common RT-PCR machines generally available in the market. The results are interpreted with the BioVisor RC software that includes the validation criteria of samples and controls.
  • The system includes an internal control of amplification based on the exclusive technology CHIC (Competitive Heterologous Internal Control) to prevent false negatives due to reaction inhibition.

What are the contents of the RealCycler CORO-G Kits v.5.3?

RealCycler CORO-G v.5.3 includes the AmpliMix CORO v.5.3 and a CORO Positive Control, which consists of a DNA artificial construction of Coronavirus. All reagents are ready to use without adding or rebuilding any component.

ComponentCORO – G VialsVolume
AmpliMix CORO v.5.3 (Retrotranscriptase included)101450 μL
CORO Positive Control1480 μL

What is the test procedure of RealCycler CORO-G Kits v.5.3?

a) Nucleic acids purification

RealCycler kits are compatible with

  • Arrow/LIAISON® IXT DiaSorin
  • BioRobot EZ1. Qiagen
  • QIAamp Viral RNA Mini kit. Qiagen
  • QIAcube. Qiagen
  • Maxwell® 16 Viral Total Purification kit. Promega Corporation
  • NucliSENS® easyMAG®. bioMérieux

 b) According to the below specifications, thermal profile Programme the “CORO” amplification protocol:

TimeTemperatureCyclesFluorescence
10:0045°C1OFF
2:0095°C1OFF
0:15
0:30
0:30
95°C
55°C
72°C
45OFF
ON
OFF

C) PCR reaction set-up

  • Thaw AmpliMix CORO v.5.3 and CORO Positive Control
  • Use as negative control RNA from negative samples, water or RNA elution buffer (not supplied).
  • Prepare the necessary amplification tubes for samples and controls (Follow the protocols as per catalogue meant for each instruments).

D) Adjust the fluorescence threshold

It is necessary to set the fluorescence threshold for each channel according to the indicated values

Key Specifications of the RealCycler CORO-G Kits

  • Certification- CE-IVD Certification ISO 9001 Certification ISO 13485 Certification
  • Other Regulatory Body-CDSCO Approved (Form MD-15) India Approved in Spain Relevant License to manufacture and import from AEMPS.
  • Sensitivity-1 copy/μL for Coronavirus SARS-CoV-2 1 copy/μL for Sarbecovirus
  • Specificity- Coronavirus SARS-CoV – ORF8 region Sarbecovirus – E gene
  • Shelf Life- stability of at least 6 months and up to 18 months
  • Store and transport samples frozen at -20 °C until use
  • Expiration date included in the external label
  • The experimental assays have shown that RealCycler reagents are stable for at least 15 cycles of freezing / thawing
  • The Reacycler kit is potentially compatible with any sample from which the pathogen’s RNA can be extracted in enough quantity and quality, although the user must assess the suitability of non-validated samples through laboratory tests.
  • Specifications BioVisor RC
    • CE mark.
    • Users and licenses management.
    • Data importation from PCR instrument.
    • Predetermined fluorescence thresholds.
    • Normalization of fluorescence values.
    • Visualization of all channels simultaneously.
    • Interpretation of the result of each sample depending of its CHIC result.
    • Series validation depending on the controls result.
    • Monitoring of possible inhibitions.
    • Allows the simultaneous quantification for several pathogens.
    • Automatic calculation of the load of pathogen in the sample.
    • Generation of a report with results.
    • Conservation of calibration curves.
    • Archiving of work series.
    • Management of all the runs from the different instruments in a single database.
    • Storage of runs obtained with the following validated instruments:
    • SmartCycler® (Cepheid®)
    • ABI 7500 (Thermo Fisher Scientific)
    • CFX96™ (Bio-Rad)
    • Mic qPCR Cycler (Bio Molecular Systems)
    • T-COR 8® (Cirrus Dx®) – Connectivity with several T-COR 8® instruments in a local area network.

CliniExperts smoothly got approval for RealCycler CORO-G Real Time PCR Kits of Progenie Molecular.

Looking for a regulatory agency to get approval of medical devices in India? You are at the right place. Call CliniExperts today for rapid and efficient approval of medical devices in India.

References

  1. Real Cycler CORO-G. Available at: https://www.progenie-molecular.com/en/assets/files/User-manualCORO-Uv.5.3.pdf .Accessed on: 9th August, 2020.