【摘要】 目的研究一种适用于副溶血性弧菌活的非可培养状态(VBNC)的检测方法。方法将副溶血性弧菌VPL4-90悬浮于陈海水中,利用低温条件诱导VBNC状态,42d后取样,进行AOAC法观察、DVC法观察、PC计数及国标法检测等;以诱导后菌体为抗原,制备兔抗VBNC血清,用于免疫捕获PCR检测。结果进入VBNC状态的细菌缩小成球形,大部分仍然是活菌,但不能形成菌落;国标检测方法未能使进入VBNC状态的细菌得到分离培养;利用抗血清捕获VBNC菌体后对副溶血性弧菌的tlh基因进行PCR扩增,检测到副溶血性弧菌VBNC。结论制备的抗血清对VBNC菌体具有富集作用;建立的免疫捕获PCR法可应用于对副溶血性弧菌VBNC的检测。更多还原

【Abstract】 Objective To study means of detecting a viable but non-culturable (VBNC) state in Vibrio parahaemolyticus.Methods The cells of Vibrio parahaemolyticus strain VPL4-90 were suspended in seawater and cultured at low temperature to induce a VBNC state.After they were cultured at low temperature for 42 days,the VBNC cells were sampled and subjected to an aerobic plate count by the AOAC method,direct count of viable bacterial cells (DVC),and plate count (PC) as well as testing by national standards.Serving as an antigen,VBNC cells were used to prepare antiserum that was then used in immunocapture-PCR detection.Results Cells in a VBNC state shrank and became spherical,and most remained alive but lost the capacity to grow into a colony.The VBNC cells could not be cultured and detected using the national standard to detect Vibrio parahaemolyticus.However,the VBNC cells were detected when captured by the anti-VBNC serum followed by PCR amplification of the tlh gene of Vibrio parahaemolyticus.Conclusion The prepared anti-VBNC serum enriched VBNC cells,and immunocapture-PCR was successfully used to detect VBNC cells of Vibrio parahaemolyticus.更多还原