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目的探讨增菌时间对沙门氏菌Real time PCR检测方法的影响。方法采集69例沙门氏菌首次检测阳性从业人员肛门拭子,置于BPW36℃孵育3、6、9和15 h,每个增菌时间段构成1个测试组,共4组。测试组样本构成组分为1份待检增菌液和9份相同增菌时长的已知阴性增菌液各50 ul。孵育15 h的时点吸取增菌液500 ul作为对照组。结果 3、6、9和15 h时长测试组阳性检出率依次为75.41%(46/61)、83.61%(51/61)、95.08%(58/61)和98.36%(60/61)。与对照组比较,经fisher检验,P值依次为0.002、0.034、0.585和1.000。结论对从业人员采用Real time PCR方法检测沙门氏菌时,扩增前的增菌时间影响到标本的检测结果,增菌时间应当保证至少9 h,15 h更为理想。
Objective To investigate the effect of enrichment time on the detection of Salmonella Real time PCR. Methods Sixty-nine salmonella samples were collected for the first time to detect the anal swabs of positive employees. The swabs were placed in BPW at 36 ℃ for 3, 6, 9 and 15 h, and each test period consisted of 4 groups. Components of the test group composed of a sample to be tested for broth and 9 copies of the same time by the known negative bacteria enrichment of 50 ul. At 15 h incubation time, 500 ul broth was added as control. Results The positive rates of the three test groups were 75.41% (46/61), 83.61% (51/61), 95.08% (58/61) and 98.36% (60/61), respectively. Compared with the control group, the P values were 0.002,0.034,0.585 and 1.000 by the Fisher test. Conclusion When using real time PCR to detect Salmonella, the pre-amplification enrichment time will affect the test results. The enrichment time should be at least 9 h and 15 h.