Objective To determine the effectiveness of Southern blot method for detection of Pla fragment in Yersinia pestis. Methods Using Southern blot and Y. pestis quantitative real-time PCR kit to detect the Pla fragment. Results The results of Southern blot were same as real-time PCR kit. Southern blot allowed to see the number of pla gene copies directly, the efficiency of hybridization was high; PCR detection was also positive. Conclusion Southern blot is suitable for detection of Pla fragment of Y. pestis.
JIN Mu-zi, YUN Hua, TAI Da-peng, CHEN Yu-fei, BAI Xiao
. Detection of Pla Fragment in Yersinia pestis by two methods[J]. Chinese Journal of Vector Biology and Control, 2017
, 28(3)
: 238
-240
.
DOI: 10.11853/j.issn.1003.8280.2017.03.010
[1] 李建云,张忠兵,范蒙光,等. 内蒙古2004年至2013年长爪沙鼠鼠疫疫源地鼠疫监测结果分析[J]. 内蒙古医学杂志, 2014, 46(9):1082-1085.
[2] 魏玉利,邱芳蕾,刘圣臣. 鼠疫菌检测方法的研究进展[J]. 生命科学仪器, 2007, 5(4):32-34.
[3] Hinnebusch J,Schwan TG. New method for plague surveillance using polymerase chain reaction to detect Yersinia pestis in fleas[J]. J Clin Microbiol, 1993, 31(6):1511-1514.
[4] Radnedge L, Gamez-Chin S,McCready PM,et al. Identification of nucleotide sequences for the specific and rapid detection of Yersinia pestis[J]. Appl Environ Microbiol,2001,67(8):3759-3762.
[5] 周长发,张锐,张晓,等. 地高辛随机引物法标记探针的Southern杂交技术优化[J]. 中国农业科技导报, 2009, 11(4):123-128.
[6] 白林庆,司晓艳,涛波. 内蒙古主要鼠疫传播媒介蚤类的分布特征及其流行病学意义[J]. 实用预防医学, 2015, 22(5):639-640,封3.
[7] Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis[J]. J Mol Biol, 1975, 98(3):503-517.
[8] Höltke HJ, Ankenbauer W,Mühlegger K,et al. The digoxigenin (DIG) system for non-radioactive labelling and detection of nucleic acids-an overview[J]. Cell Mol Biol, 1995, 41(7):883-905.
[9] McCabe MS,Power JB, de Laat AMM,et al. Detection of single-copy genes in DNA from transgenic plants by nonradioactive Southern blot analysis[J]. Mol Biotechnol, 1997, 7(1):79-84.
[10] Matthews JA,Kricka LJ. Analytical strategies for the use of DNA probes[J]. Anal Biochem, 1988, 169(1):1-25.
[11] 龚正达,于心,刘起勇,等. 中国鼠疫自然疫源地分型研究Ⅵ. 鼠疫媒介生物学特征[J]. 中华流行病学杂志, 2012, 33(8):818-822.
[12] 秦长育,许磊,张荣祖,等. 中国鼠疫自然疫源地分型研究Ⅴ. 鼠疫宿主生物学特征[J]. 中华流行病学杂志, 2012, 33(7):692-697.
