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目的研制适用于副溶血性弧菌(Vibrio Parahaemolyticus,VP)能力验证(PT)项目的 VP菌及干扰菌的冷冻干燥样品,建立有效的样品制备与评估流程。方法通过不同比例混匀的VP和干扰菌拟态弧菌(Vibrio minicus,VM)经冻干后在鉴定培养基上菌落生长优势比,确定制备PT样品的混合菌VP和VM的浓度比例;通过系统抽样和鉴定以评估样品的均一性;通过观察样品在180 d保存期内菌量的变化情况以评估样品的稳定性;根据188间检测实验室反馈的鉴定结果以评估和计算样品的实际合格率。结果选择5 000∶1比例制备的VP和VM混合PT样品能在鉴定培养基有效分离两种菌落;抽检的40份样品均能有效鉴定,显示PT样品具有较好的一致性;通过保存温度和时间的稳定性实验显示,PT样品在36℃下能稳定保存5 d,冻干后PT样品于4℃保存180 d后VP菌仍能检出;188家参与实验室检测结果显示,PT样品合格率为97.3%。结论本研究制备的冻干PT样品及实验流程控制适用于副溶血性弧菌及类似的微生物鉴定能力验证项目。
OBJECTIVE To develop freeze-dried samples of Vibrio parahaemolyticus (VP) proficiency test (VP) and interfering bacteria and to establish an effective sample preparation and evaluation procedure. Methods Vibrio minicus (VM) mixed with VP and Vibrio minicus (VM) in different proportions were used to determine the concentration ratio of VP and VM in the mixed culture of PT samples. Sampling and identification to assess sample homogeneity; assessing sample stability by observing changes in bacterial volume over a 180-day shelf life; assessing and calculating the actual pass rate of samples based on the 188 lab feedback laboratory qualification results . Results The VP and VM mixed PT samples prepared at the ratio of 5: 1 were able to effectively separate the two colonies in the identification medium. All the 40 samples were validated and showed that PT samples had good consistency. The stability of time experiments showed that the PT samples can be stably stored at 36 ℃ for 5 days and the samples of the VP samples can still be detected after the samples were lyophilized for 180 days at 4 ℃. The results of 188 laboratory tests showed that the PT samples were qualified The rate was 97.3%. Conclusion The freeze-dried PT samples prepared in this study and experimental flow control are suitable for Vibrio parahaemolyticus and similar microorganism identification proficiency testing programs.