Quality control

1.    Scope:

·       This SOP explains the quality control measures adopted in the polio laboratory to ensure correct and reproducible results. Quality control measures are performed in every compartment of the laboratory.

2. Procedure:

Internal Quality Control

External Quality Control

Internal Quality Control Measures for Cell Line

·       Sterility of every batch of prepared media and reagents like FBS is checked for 7days and entered in file No.PF 5. (Refer SOP 6.3)

·       Titration of both cell lines (L20 B and RD) is done at 3^rd and 8th passage with laboratory quality control standards and readings are compared with standard reference values for each of the polio virus type. Refer SOP 6.1

·       Each cell line is inspected macroscopically and microscopically upon receiving in the laboratory. Its titration is performed after splitting.

·       LN2 level is checked every 15 days and whenever it is opened.

Internal Quality Control for Virus Isolation

·       A panel of stool samples is prepared in the laboratory which is used to carry out the internal quality assessment for virus isolation. Every 3 months 10 samples are picked by lab incharge and given for processing for virus solation and identification including ITD.

Internal Quality Control for rRTPCR for ITD and VDPV

·       With every run a negative control and a positive control for each primers (Pan EV, Sabin123, Pan PV, Sero123, and VDPV S123) is tested.

External Quality Control for virus isolation

·       For virus isolation, once a year, 10 samples are received from WHO head office. Samples are processed as per protocol and reported to Virologist ramamurtyn@searo.who.int and harrie.van.avoort@rivm.nl

·        Results are submitted within 14 days.

External Quality Control for ITD

·       Once a year 10 samples are received from WHO for ITD and VDPV.

·       Results are submitted in 7 days to Mmandelbaum@cdc.gov and Virologist WHO.

Other quality control measures

Sterility testing of Bio Safety Cabinets: once in a month

Procedure

·       Clean the biosafety cabinet with 1% Hypochlorite solution before any work in the morning.

·       Keep U.V. light and air flow on for 10min.

·       Label five N –Agar plate labeled with hood No. and position placed in the BSC. (2nLeft, 2 right, middle).

·       Switch off U.V. light and take three previously labeled N – Agar plates.

·       Place it in the BSC according to the label and open lid of N – Agar plate. Close BSCs and keep it for 5 min.

·       After 5 min. close the lid of N – Agar plate and put it in incubator at 360C.

·       Result after 24hrs.and record it in sterility testing PR 41

·       Temperature of incubators, refrigerators and deep freezers are noted in temperature register and marked on graphs every morning and evening.

·       Incubators are cleaned with 70% alcohol once in a week and recorded in Maintenance of Incubators File No. PF-19

·       Deep freezers are defrosted and cleaned once in a months and recorded in their respective log books.

·       Titration of cell culture done 3 ^rd and 8^th passage, when there is any change in cell culture like new lot of media, new lot of serum, new technician, new incubator or any other major change titration is done.

·       Autoclave and oven cycle noted regularly. Autoclave tape is used as an indicator to check proper Autoclave cycle. It is also checked with biological indicator once in a month.

·       Microscope service is done regularly.

·       In Thermocycler 7500 we do all calibreations Back ground calibration,Optical calibration, ROI calibration & Dye calibration after every six months.

    3. Documentation

·       Interenal Quality Control Register PR-15

·       Interenal Quality Control File PF-14