样本检前保存条件对幽门螺杆菌培养结果影响

Effect of sample preservation conditions on the culture result of Helicobacter pylori

目的 研究幽门螺杆菌样本培养前贮存温度、贮存时间和运输方法对结果影响。方法 将标准幽门螺杆菌菌株置运输培养基中放置于4、25、35、-25、-80 ℃环境中,分别存放6.0、12.0、24.0、48.0 h后进行微需氧培养。将标本置于4 ℃冷链运输箱,通过汽车、火车和飞机运输,并且在存放6.0 h接种培养,同时以 0.0 h 样本为对照组。结果 固体培养到第3天时幽门螺杆菌生长良好,哥伦比亚血琼脂平板上见透明、针尖样菌落形态;幽门螺杆菌为革兰阴性的螺旋状、海鸥状、s状弯曲菌或短杆菌。样本贮存温度与存放时间对结果有非常显著性影响。样本贮存24.0 h内,以贮存4 ℃环境样本阳性率最高(40.4%±5.1%),与其他各组比较差异有统计学意义( P <0.05);在贮存6.0 h以内,以贮存25 ℃阳性率最高(71.2%±8.1%),与35、-25、-80 ℃比较,差异有统计学意义( P <0.05)。幽门螺杆菌样本经过汽车(幽门螺杆菌存活率为50.7%± 5.8 %)、高铁(幽门螺杆菌存活率为52.1%±6.7%)、飞机(幽门螺杆菌存活率为3.2%±3.3%)运输后6.0 h再行培养以飞机运输阳性率最低,与其他运输方式结果比较,差异有统计学意义( P <0.05)。结论 样本贮存时间和温度对幽门螺杆菌培养阳性率有明显影响,通过飞机运输可明显降低幽门螺杆菌培养的阳性率。

Objective To study the effects of storage temperature,storage time and transportation methods on the results of Helicobacter pylori samples before culture. Methods The standard strains of Helicobacter pylori were placed in transport medium at 4,25,35,-25 and -80 ℃ for 6.0,12.0,24.0 and 48.0 hours,respectively.The specimens were transported by car,train and airplane in a cold chain container at 4 ℃ and inoculated for 6.0 h.Meanwhile,the 0.0 h specimens were used as control group. Results Helicobacter pylori grew well on the 3rd day of solid culture.Colombian blood agar plate showed clear,needle-tip-like colonies.Helicobacter pylori was Gram-negative spiral,gull-shaped,Campylobacter S-shaped or brevis.Sample storage temperature and storage time have a very significant impact on the results.Within 24.0 hours of storage,the positive rate of environmental samples stored at 4 ℃ was the highest (40.4%±5.1%),which was significantly different from other groups ( P <0.05).Within 6.0 hours of storage,the highest positive rate was ( 71.2%± 8.1%) at 25 ℃,compared with 35,-25,-80 ℃,the difference was significant ( P <0.05).Helicobacter pylori samples were cultured in automobiles (50.7%±5.8%),high iron (52.1%±6.7%) and airplanes ( 3.2%± 0.3%) at 6.0 hours after transportation,the positive rate of Helicobacter pylori samples was the lowest in airplane transportation,compared with the results of other transportation modes,the difference was statistically significant( P <0.05). Conclusion Sample storage time and temperature have a significant impact on the positive rate of Helicobacter pylori culture.Airplane transportation can significantly reduce the positive rate of Helicobacter pylori culture.