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What to do if there is contamination in the PCR laboratory
2007-01-16 17:19
When it is found that the PCR laboratory is contaminated, we first consider that it may be the contamination of the reagent. After re-testing with a new batch of reagent, the result is still positive for all specimens.
Take the specimen to other formal PCR laboratories for testing. The result has negative and positive results. The negative control is negative and the positive control is positive, indicating that the specimen is not contaminated. Take two batches of reagent to other regular PCR Laboratory tests also showed that there was no problem with the reagents.
After wiping the work surface with disinfectant in our laboratory, turn on all the UV lamps for one day, replace the micro sampler, centrifuge tube, gun head, gloves, marker, record book, etc., and then carry out the specimen. Test, all specimens are still positive.
After wiping the work surface with disinfectant, turn on all the UV lamps and irradiate for one day to re-examine the specimen. At the same time, two negative controls (A and B) are set. A negative control is to take the reaction tube of the reagent from the reagent box. Open the lid of the reaction tube, expose it for 10 min in the air, close the lid, and test it on the machine; B negative control is to take the reaction tube of the reagent from the reagent box without opening the lid, and directly test it on the machine. The result is that all specimens and A negative control are positive, while B negative control is negative.
So far, although it is still not possible to find out what the source of the pollution is, one thing is for sure. The PCR laboratory pollution was caused by the aerosol diffusion. After investigation, it was found that the pollution was due to a worker in the laboratory cleaning the laboratory. Hygiene caused by sweeping the swab and swab in the amplification area to clean the specimen preparation area.
After determining the cause of the pollution, we began to eliminate the pollution. Every day, we opened the doors and windows of the entire PCR laboratory and the exhaust fan to ventilate the entire laboratory, wiped the entire PCR laboratory with disinfectant, and turned on all UV lamps. .
According to the step 3.5 test every 3 days, it has been treated in this way for one month before the A negative control turns negative. The A and B negative controls were tested three times in succession, and each result was negative to determine that the PCR laboratory returned to normal.
Generally, the prevention of PCR laboratory pollution is to isolate the different operation areas, repack reagents, and improve the experimental operation rules. The treatment of pollution is to use dilute hydrochloric acid to wipe or soak; ultraviolet irradiation method. However, enough attention has not been paid to the ventilation of the laboratory. In order to prevent the mutual contamination between the experimental sections, each experimental section is sealed as much as possible, thereby increasing the chance of residual contamination of PCR products in the laboratory.
Judging from the process of contamination elimination in this PCR laboratory. I think one of the keys to pollution is ventilation. Properly giving laboratory ventilation can play the role of air dilution, which is beneficial to the reduction of PCR product residues and speeds up the elimination of laboratory pollution.

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