Candle jar fractional cultivation and cryopreservation of Helicobacter pylori

doi:10.3969/j.issn.1008-0171.2011.05.020

Abstract：Objects To study the simple method for the separation and reservation of Helicobacter pylori（H pylori） and construct a small storage of H pylori.Method Firstly,the candle jar method was used to separate samples of H pylori from different sources by H pylori selective medium,Secondarily,the Gram microscopy test and biochemical experiment were used to identify H pylori.Thirdly,the pure H pylori was reserved in three different preservation solutions（brucella broth,50% glycerol serum,50% glycerol egg white） which were 20 °C below zero.Finally,the samples were tested to find out the survival rate after 1-,6-,12-day reservation.Result Among the 21 H pylori samples,of which 13 were collected and obtained from 58 clinical specimens and the other 8 were from oral and fecal samples of 56 undergraduates,the survival rate of 21 of the samples was over 90%.That is,the survival rate showed no significant difference when they were tested after 1-and 6-day reservation.The survival rate of the samples reserved in different solutions showed significant different when tested in 12-day reservation： that of those in 50% glycerol serum was over 80%,that of those in the other solutions was less than 50%.Conclusion Candle jar method can be used as a good way to isolated H pylori and the 50% glycerol which is 20 °C below zero can reserve H pylori well for more than 12 days.It is convenient to construct a simple and small storage of H pylori and valuable in scientific research by using the bacteria from the storage.