NABL launched online portal for "NABL Accreditation for medical testing laboratories of RT-PCR RNA Virus/ COVID-19". Accreditation was being provided within 10 days of sending online application through online assessment (through Zoom app). Lab needs to get NABL Accredited for any one Infectious RNA Virus - RT PCR Tests (Such as HCV, HIV, Dengue).
Hereby provided basic layout without measurement for molecular testing Lab for RT-PCR Testing of RNA Virus.
Below provided layout can be used as basic reference for designing the molecular testing lab for medical diagnostics.
Note : For TrueNat or Catridge based PCR Methods, PCR Master mix & Template Addition rooms may not required. However contact the manufacturer representatives before start of infrastructure work for PCR Lab.
Note : ICMR Providing Expedited Approval ( within 7 days) for Covid 19 Testing based on submitting Lab Details and NABL Application copy For TrueNat or CBNAAT PCR Equipment's. Lab should get NABL Accredited and Scope & Accreditation Certificate copy should be submitted to ICMR within 4 weeks from the date of Provisional Approval for Covid 19 RT PCR Testing.
ICMR Approval Requirements for Setting up a Real Time PCR Testing Facility for Covid -19
Staff , Equipment and Consumables Requirements
(Excerpts from "ICMR - Application invited from Govt & Private Medical Colleges for setting up Covid-19 testing facility, date 8.4.2020")
Staff Requirements :
A. Availability of following minimum staff:
Medical Microbiologists – 1 or more with experience of work in Molecular Virology.
Technicians – At least 4-6 (2-3/shift) with relevant experience of work in Molecular Virology.
Multi-Task Staff – 1 or more for washing / cleaning
B. Desired expertise of the staff:
Good understanding of laboratory biosafety and biosecurity, trained for handling respiratory samples for viral diagnosis, RNA extraction and real-time PCR.
Experience of work in virology and handling clinical specimens, especially respiratory samples.
Equipment's :
- Biosafety cabinet (BSC) class 2A or 2B2 (calibrated)
- -20 °C deep freezer with UPS, for storage of reagents (primers/ probes/ positive controls)
- -80°C deep freezer with UPS, for storage of aliquoted samples/ viral RNA in cryovials
- 4°C refrigerator (for storage of viral transport medium, and for short term storage of samples and extracted RNA)
- UPS (2 nos., 2KVA each, with 2 hours back-up, for real time PCR instrument and nucleic acid extraction systems – if not available, then to be carried); and confirm about power backup for the two deep freezers (check about duration of power outages, if any)
- Real-time PCR machine
- Microcentrifuge / Refrigerated Centrifuge
Consumables :
- For sample collection:
a. Personal protective equipment (PPE)
b. Viral Transport Medium (VTM)
c. Flocked Dacron swabs (2 swabs/ sample collection from 1 patient
- During Processing:
a. Biosafety cabinet class IIA/ IIB
b. Personal protective equipment: N95 masks, coveralls (protective against blood and body fluids), nitrile gloves, shoe cover, head cover
c. Vortex mixer
d. Microcentrifuge (Cold centrifuge)
e. Cryovials (2 ml)
f. Cryobox
g.Pipette aid
h.Disposable plastic pipettes
i. Sprit lamp
j. Forceps (if no spirit lamp, then disposable forceps for each sample)
k. 70 % ethanol (also required for next stage, i.e., extraction)
l. 1% sodium hypochlorite (4% stock, to be freshly reconstituted daily to 1% with water)
m. Discarding jars
n. Biomedical waste disposal (BMW) bags (with ties for sealing; preferably autoclavable, if discarding autoclave is available/ used locally) and bins
o. Iceboxes with gel packs or regular ice supply in laboratory (from icemaker)
p. Tube rack (15 ml tubes)
q. For tube / cryovial labelling - Marker pens, cellotape, or label printouts (printer with label maker)
Nucleic Acid Extraction
- Manual extraction using kits for Viral RNA extraction: Viral RNA mini kits (Qiagen) or other viral RNA extraction kits for manual extraction
- 1.7 ml Eppendorf tubes (separate ones also required for next step)
- Cryovial/ Eppendorf tube rack (separate ones also required for next step)
- Microcentrifuge (small equipment)
- Micropipettes- 100-1000ul, 20-200ul (additional separate micropipettes of required volumes also listed for next stage, i.e. real time PCR)
- Filter barrier tips: 1000ul, 200ul
- Tissue rolls
- Hand sanitizers
- Biohazard labels
Real Time PCR
- Real time PCR machine (open system) – calibrated for the fluorophore dyes which are present on the probes .
- Reagents for setting up Real-time
PCR primers and probes specific for SARS-CoV2 targets
PCR master mix reagents (e.g., Thermo Fisher/ Invitrogen AgPath/ Superscript III Platinum real time PCR reagents) with buffer and enzyme
- PCR Reagents
Primers for E gene screening and
Probes for E gene screening and RDRP/ ORF 1b targets
PCR Buffer and enzyme mix o Positive contro
- PCR workstations – 1 for mastermix preparation; 1 for RNA addition
- Cryovial racks
- PCR tubes/ PCR plates
- PCR plate adhesive seals and plate sealer
- Micropipettes- 0.5-10ul (2 nos., 1 for PCR master mix and 1 for RNA addition), 2-20ul,20-200ul
- Filter barrier tips – 10ul, 20ul, 200ul
- Microspin (small equipment)
- Plate centrifuge (small equipment)
- Electronic micropipette (optional small equipment, but convenient and reduces time duration of testing)
- Nuclease free water - for PCR
- RNaseP
NABL Guidelines for Storage of Molecular Testing Specimens :
(NABL 112 Guideline, Issue 4, Issue Date 11.2.2019)
|
Test/Sample |
Minimum Retention Period & Storage Conditions |
|
DNA |
5 Years at -20°C 2 Years at -20°C for infectious disease molecular Testing |
|
Extracted RNA |
5 Years at -70°C 2 Years at -70°C for infectious disease molecular Testing |
Source Reference :
1. https://www.icmr.gov.in/ctestlab.html
2. https://nabl-india.org/
3. https://apps.who.int/iris/handle/10665/249549
POSTED BY:
Mr.V.SELVAM, M.Sc (Biochemistry)
Consultant - ISO 9001 & NABL (ISO 15189 & ISO 17025)
Chennai, Tamilnadu, India
✆ +91-9940014466
