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BARC

A CRISPR-based integrated system for detection of Covid-19

An ultra-portable system for carrying out CRISPR based Covid-19 detection reaction that includes a module for isothermal amplification (RT-LAMP reaction) followed by CRISPR based detection of the viral RNA. The results are directly visualized with the integrated Fluorescence module. The kit detects the N2 and E fragment of SARS-CoV-2 virus. RNA extracted from patient is converted to DNA and amplified using RT-LAMP. The amplified DNA is detected by CRISPR-Cas12-gRNA-complex. This triggers cleavage of a DNA probe resulting in emission of fluorescence. The increase in fluorescence is indicative of a positive reaction. The kit uses RNAseP as an internal control.

An ultra-portable system for carrying out CRISPR based Covid-19 detection reaction that includes a module for isothermal amplification (RT-LAMP reaction) followed by CRISPR based detection of the viral RNA. The results are directly visualized with the integrated Fluorescence module. The kit detects the N2 and E fragment of SARS-CoV-2 virus. RNA extracted from patient is converted to DNA and amplified using RT-LAMP. The amplified DNA is detected by CRISPR-Cas12-gRNA-complex. This triggers cleavage of a DNA probe resulting in emission of fluorescence. The increase in fluorescence is indicative of a positive reaction. The kit uses RNAseP as an internal control.

Salient Features
  1. Isothermal amplification and CRISPR based detection
  2. Compact (10 cm cube), Battery operated
  3. Microprocessor controlled - Programmable temperature and timer controls
  4. Module for visualization of the end-product (Visual / Electronic read out)
  5. Single push button operation and LED indicators for reaction progress
  6. Can analyse five samples in each run.
  7. Does not need any additional instrumentation.
  8. The method evaluated using more than 100 (synthetic RNA as well as clinical) samples.
  9. System stability and performance assessed in more than 25 runs.
  10. This technology is ready to be taken forward for field testing, ICMR approval and deployment.

An ultra-portable system for carrying out CRISPR based Covid-19 detection reaction that includes a module for isothermal amplification (RT-LAMP reaction) followed by CRISPR based detection of the viral RNA. The results are directly visualized with the integrated Fluorescence module. The kit detects the N2 and E fragment of SARS-CoV-2 virus. RNA extracted from patient is converted to DNA and amplified using RT-LAMP. The amplified DNA is recognized by CRISPR-Cas12-gRNA-complex and detected by fluorescence. The kit uses RNAseP as an internal control.

Salient Features
  1. Isothermal amplification and CRISPR based detection
  2. Compact (10 cm cube), Battery operated
  3. Microprocessor controlled - Programmable temperature and timer controls
  4. Module for visualization of the end-product (Visual / Electronic read out)
  5. Single push button operation and LED indicators for reaction progress
  6. Can analyse five samples in each run.
  7. Does not need any additional instrumentation.
Applications
  1. For diagnosis of infection with coronavirus 2 (SARS-CoV-2) causing Covid-19 disease
  2. Principle can be extended to other similar infections
Specifications

Kit components

  1. Time for detection: 75 minutes
  2. Limit of detection: 200 viral RNA copies
  3. Simultaneous detection of N2 and E genes

Instrument

  1. Temperature Range: Ambient to 70°C
  2. Sample Holder: 16 tubes of 0.2 ml each (PCR tubes)
  3. Temperature accuracy: ± 0.5 °C at the set point
  4. Timer Control: in steps of 1 second each up to 99 hours 
  5. Mode of Control: PID
  6. Indicators: Blinking LED to indicate the progress
  7. Audio Feed back: Buzzer
  8. System Dimensions: 10 cm X 10 cm X 10 cm
  9. Connectivity: USB and Bluetooth
  10. Power Supply: 5 V, 2 A adopter or a rechargeable battery of 10,000 mAH.
INFRASTRUCTURE REQUIREMENT

Machine shop with milling machine, electrical circuit assembly and testing facility. Biological material handling facility.

Space

  1. Instrument foot print            :           100 sq. cm
  2. Total working space            :           50 sq. meter

Raw Material

  1. Most of the components are available within the country and some components (Cas12, LAMP mix.) need to be imported through local agents.

Manpower

  1. One graduate in science, one electronics engineer, two trained technicians will be required for manufacture/production of this instrument.
Vendors' details for various components and materials

Sr. No. Items Vendor details
1 Mechanical components M/s Bright Engg Works, Balaji Industrial Complex,
F-5,Tonadr Village , Near - Hindlco ltd, Taloja MIDC, Panvel. Dis: Raighad. PIN- 410208
Phone: 9167555999 / 9869329259 (brighteng85@gmail.com)
2 Electronics 1. M/s Dyptronics, Mumbai
2. M/s Brilliant electronics, Mumbai
3 Biochemicals M/s Zelle Biotechnology Pvt Ltd,
A7 MIDC, Mira Industrial Area, Western Express Highway, Mira Road, Thane-401104.
info@zellebiotech.com
Application Procedure
BARC Technologies