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Objective:To observe the stability and sensitivity of transcription mediated ampliifcation (TMA) system, and to compare it with real-time reverse transcription polymerase chain reaction (RT-PCR) in amplifying serum HCV RNA in HCV infected patients. Methods: TMA system was established by moloney murine leukemia virus (MMLV) reverse transcriptase, T7 RNA polymerase and 2 speciifc primers ifrstly,and then its stability and repeatability were compared at different storage temperatures by the correlation change of HCV RNA amplification curve. The sensitivity difference between TMA and RT-PCR was evaluated by amplifying a group of 10-fold diluted HCV RNA samples which were transcribed in vitro. A total of 101 serums of chronic HCV infected patients were measured by TMA system and RT-PCR to observe the positive rate and their correlation. Linear correlation and linear regression were used to observe the correlation of the two methods. Results:TMA system was successfully established. TMA system was not stable when stored at 20℃ (placed for 24 hours only), but it was stable for 6 days when stored at 4℃ or within 6 months when stored at-20 ℃. Compared with RT-PCR whose reagent was made by Hunan Sansure Biotechology Corporation, TMA system showed 20 positive samples and 11 negative samples in a total of 31 samples. So was the RT-PCR kit of the Sansure Biotechology Corporation, and the concordance rate of the two methods was 100%. Advanced quantitative study of the 20 positive samples found that the two methods had good correlation and consistency (r=0.91,P0.05). Advanced quantitative study of 29 positive samples found that the two methods had good correlation and consistency (r=0.96,P<0.01). Conclusion:The stability and repeatability of TMA system are good within 6 months when stored at-20 ℃ storage temperature. Both TMA and RT-PCR HCV RNA can detect serum HCV RNA well, and the two methods have good correlation and consistency.
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OBJECTIVE To compare the effect with the three methods to disinfect the ventilator circuits.(METHODS) To disinfect the ventilator circuits,three methods were used:A,soaking with chlorine compoud(disinfectant);B:soaking with 0.5% peracetic acid;C,sterilizing with ethylene oxide.Samples were taken to(bacteria) culture before and after disifection in every three methods.Biological supervision and the amount of colony were also taken at the same time.RESULTS The results of bacteria culture pre-disinfection were all positive.The amount of bacteria of ventilator circuits was between 70 000-120 000CFU/cm~2.The eligible rate of biological(supervision) in method C was 100% and its positive rate of bacteria culture was 0%.The eligible rate of biological supervision in method A was 86.25% and its positive rate of bacteria culture was 16.25%.The eligible rate of(biological) supervision in method B was 90% and its positive rate of bacteria culture was 13.78%.It was significant when method C was compared with methods A and B and the value P was less than 0.01.CONCLUSIONS It is(serious) about the pollution in ventilator circuits.The soaking methods are apt to be effected by the factors of(environment) and manual operation.Therefore it could be(ineligible) and being polluted after disinfection.The method of sterilization is the best for disinfection.
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Objective To find out sterilization quality of pulsating vacuum sterilizer for better sterilization effect.Methods The sterilization quality was monitored by physical,chemical and biological methods.Results 4,576 subjects were involved in the test.Such data were obtained as the qualification rate for temperature,the one by chemical indicator card,the one by B-D test and the one by biological indicator,which were 99.6%,99.8%,99.9% and 100% respectively.Conclusion Above data proves that the sterilizer is in normal condition.