发光细菌DH132的筛选、鉴定及其在氯霉素生物毒性测定中的应用  

Screening and identification of luminescent bacteria and its application in chloramphenicol detection

作　　者：杜宗军[1] 林瀚智[1] 王静雪[1] 池振明[1] 吴龙飞[1] 

机构地区：[1]中国海洋大学生命科学与技术学部,青岛266003

出　　处：《高技术通讯》2007年第12期1290-1294,共5页Chinese High Technology Letters

基　　金：国家自然科学基金(30500384);中国海洋大学"本科生训练计划"(0611010404)资助项目

摘　　要：利用药敏纸片法,从本实验室保存的发光细菌中筛选出一株对氯霉素敏感的细菌DH132。通过生理生化特征鉴定并利用16SrDNA序列分析的方法,鉴定细菌DH132为哈维氏弧菌。通过在细菌培养液中添加不同浓度的氯霉素溶液,研究了氯霉素浓度与细菌发光强度抑制率之间的关系。结果表明,当氯霉素浓度为80ng/mL和160ng/mL时,对细菌发光的抑制率分别达到10%和50%。氯霉素浓度在0~60ng/mL,60~140ng/mL,140ng/mL^200ng/mL和200~1000ng/mL之间,细菌发光抑制率与氯霉素浓度分别具有很好的线性关系。相对应的回归方程分别为y=0.0627x-0.0233;y=0.3599x-18.276;y=0.7527x-70.219;y=0.0195x+75.964,回归曲线R2值分别为0.9805,0.9918,0.9502和0.9731。A marine bioluminescent bacterium, strain DH132, was selected as the test organism in chloramphenicol toxicity test. On the basis of biochemical and 16S rDNA molecular characteristics, this strain was identified as Vibrio harveyi strain DH132. The 2dh bioluminescence inhibition assay using strain DH132 was carried out to determine the relationships between the concentration of chloramphenicol and the luminous intensity of the bacteria. The standard 10% and 50% inhibition concentration （ECl0 EC50） for chloramphenicol were 80ng/mL and 160ng/mL, respectively. Four linear concentration-inhibition relationships were observed in the regions of 0 - 60ng/mL, 60 - 14Ong/mL, 14Ong/mL 200ng/mL and 200 ~ 1000ng/mL, respectively. Four curves of concentration-inhibition of chloramphenicol were obtained when the concentrations of chloramphenicol were in those regions. The regression equations were： y = 0.0627x - 0.0233, y = 0.3599x - 18.276, y = 0.7527x - 70.219, y = 0.0195x ＋ 75.964, respectively. And their correlation co- efficient （R square） of regression curves were 0.9805, 0.9918, 0.9502 and 0.9731, respectively.

关 键 词：氯霉素 发光细菌 哈维氏弧菌 抑制 毒性测定 

分 类 号：Q93[生物学—微生物学]