Detection and identification of Bacillus cereus susceptible to phage AP631
ZHANG Hui-juan1, LIU Dong-li2, HE Li3, LI Wei1, ZHANG En-min1, WEI Jian-chun1, JI Jian-jun3,XU Su-li3, LIU Wei3, ZHANG Fa-xin3, MA Guo-zhu2, SHI Yi2, MA Lin2, LIANG Xu-dong1
1. National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention, Beijing 102206, China; 2. Shanxi Provincial Center for Disease Control and Prevention, Xi’an 710054, China; 3. Yan’an Center for Disease Control and Prevention, Yan’an 716000, China
Abstract:We identified the isolates which were susceptible to phage AP631, a specific Bacillus anthracis susceptible phage. Five isolates of Bacillus cereus from soil in an anthrax endemic area and 3 preserved in our laboratory were identified by regular experiments including penicillin inhibition experiment, smear Gram staining, hemolysis testing, biochemical reaction, PCR analysis, mouse toxicity determination, protein crystallization dying experiment and MLST subtyping. All the tests supported the new bacterial isolates susceptible to AP631 phage was B. cereus and excluded B. anthracis and B. thuringiensis. Three laboratory preserved isolates were B. cereus which were identified previously. The five B. cereus isolates from soil were resistant to penicillin and PCR amplification targeted virulence genes showed negative in these strains. Protein crystallization dying experiment also displayed negative results, but the resultant biochemical reactions confirmed the 5 strains were B. cereus. In addition, all 8 B. cereus strains were clustered as new allelic loci by MLST subtyping method. Our results demonstrate that bacterial phage AP631 is capable of lysing B. cereus in addition to B. anthracis, which suggests that there is some nonspecific lysis produced by AP631 and thereby we need pay more attention to the diagnosis of B. cereus infection to avoid misdiagnosis.
张慧娟, 刘东立, 贺莉, 李伟,张恩民,魏建春,纪建军,徐苏丽,刘玮, 张发信,马国柱,石一,马琳,梁旭东. AP631炭疽噬菌体裂解蜡样芽胞杆菌的发现和鉴定[J]. 中国人兽共患病学报, 2016, 32(6): 507-511.
ZHANG Hui-juan, LIU Dong-li, HE Li, LI Wei, ZHANG En-min, WEI Jian-chun, JI Jian-jun,XU Su-li, LIU Wei, ZHANG Fa-xin, MA Guo-zhu, SHI Yi, MA Lin, LIANG Xu-dong. Detection and identification of Bacillus cereus susceptible to phage AP631. Chinese Journal of Zoonoses, 2016, 32(6): 507-511.
[1] Liang XD. A Manual of Anthrax Control and Treatment.[M]. Beijing: China Agricultural Press, 1995:27-30. (in Chinese)梁旭东.炭疽防治手册[M].北京:中国农业出版社,1995:27-30. [2] Wang BX, Zhang SR, Yu XM, et al.Characteristics of Bacillus anthracis bacteriophage AP631[J]. Chin J Virol, 1992, 8(2): 14-17. (in Chinese)王秉翔,张守让,虞雪梅,等.炭疽芽胞杆菌噬菌体AP631的生物学特性检测[J].病毒学报,1992,8(2):14-17. [3] Dong SL, Wang BX. Anthrax[M]. Xi’an: Shanxi Science and Technology Press, 2004:79-87. (in Chinese)董树林,王秉翔.炭疽[M].西安:陕西科学技术出版社,2004:79-87. [4] Davison S, Couture-Tosi E, Candela T, et al. Identification of the Bacillus anthracis(gamma) phage receptor[J]. J Bacteriol,2005,187(19):6742-6749. [5] Ash C, Farrow JA, Dorsch M, et al. Comparative analysis of Bacillus anthracis,Bacillus cereus, and related species on the basis of reverse transcriptase sequencing of 16S rRNA[J]. Int J Syst Bacteriol,1991,41(3):343-346.