微滴单细胞培养技术应用于白色念珠菌培养的研究
|
摘要
目的为提高白念菌感染的检出率及建立微滴单细胞培养检测方法提供依据。方法将白色念珠菌标准菌株分别接种在4种不同培养基上培养,将临床标本加入到含青霉素和链霉素(双抗)的培养基中进行微滴培养。结果白色念珠菌标准菌株在营养肉汤培养基中生长最快。临床样本在含双抗的培养基的生长速度6 h后快于对照组,差异有统计学意义(P<0.05)。临床样本经过18 h的微滴单细胞培养和液态培养,细胞数量大于0 h,差异有统计学意义(P<0.05)。结论临床样品在含双抗的营养肉汤培养基微滴单细胞培养18 h后再进行检测能够提高白色念珠菌感染的检出率。
Objective To improve the detection rare of Candida albicans infection and offer support for micro-drop singlecell culture detection.Methods The standard strains of Candida albicans were inoculated on four different culture media,and the clinical samples were added to the medium containing penicillin and streptomycin(double antibodies)for microtiter culture.Results The standard strain of Candida albicans grew fastest in the nutrient broth medium.The clinical samples grew significantly faster than those in the control group after 6 h(P<0.05).The number of cells was significantly greater than0 h after 18 h microcell culture or liquid culture(P<0.05).Conclusion The detection rate of Candida albicans infection could be improved by the clinical samples after single-cell culture for 18 h in a micro-droplet culture medium containing double-antibiotics.
Objective To improve the detection rare of Candida albicans infection and offer support for micro-drop singlecell culture detection.Methods The standard strains of Candida albicans were inoculated on four different culture media,and the clinical samples were added to the medium containing penicillin and streptomycin(double antibodies)for microtiter culture.Results The standard strain of Candida albicans grew fastest in the nutrient broth medium.The clinical samples grew significantly faster than those in the control group after 6 h(P<0.05).The number of cells was significantly greater than0 h after 18 h microcell culture or liquid culture(P<0.05).Conclusion The detection rate of Candida albicans infection could be improved by the clinical samples after single-cell culture for 18 h in a micro-droplet culture medium containing double-antibiotics.
引文
[1]冯兆民,舒跃龙.数字PCR技术及其应用进展[J].病毒学报,2017,33(1):103-107.Feng ZM,Shu YL.An overview of digital PCR[J].Chinese Journal of Virology,2017,33(1):103-107.
[2]蒋析文,朱小亚,高秀洁,等.数字PCR在几种常见人类传染病研究中的应用[J].分子诊断与治疗杂志,2017,9(1):50-54.Jiang XW,Zhu XY,Gao XJ,et al.The applications of digital PCR on the researches of several common human infections[J].J Mol Diagn Ther,2017,9(1):50-54.
[3]赵新,兰青阔,陈锐,等.应用微滴数字PCR技术快速检测食用菌中沙门氏菌[J].食品与生物技术学报,2017,36(3):315-321.Zhao X,Lan QK,Chen R,et al.Rapid detection of Salmonellla spp.in edible fungi by droplet digital PCR[J].Journal of Food Science and Biotechnology,2017,36(3):315-321.
[4]丰蕾,杨帆,李彩霞,等.单细胞分离检验技术在法医学中的应用研究[J].刑事技术,2015,40(5):359-363.Feng L,Yang F,Li CX,et al.Single cell separation testing and its forensic application[J].Forensic Science and Technology,2015,40(5):359-363.
[5]姚金光,解继胜,李俊,等.舌癌单细胞培养建系与癌干细胞相关标志的检测[J].华西口腔医学杂志,2013,31(1):86-90.Yao JG,Xie JS,Li J,et al.Establishment of subseries cell lines from tongue cancer single cell and detection of cancer stem cell markers[J].West China Journal of Stomatology,2013,31(1):86-90.
[6]占爱瑶,由香玲,詹亚光,等.白桦单细胞培养体系的建立[J].植物生理学通讯,2010,46(4):370-374.Zhan AY,You XL,Zhan YG,et al.Establishment of single cell culture of white birch(betula platyphylla suk)[J].Plant Physiology Communications,2010,46(4):370-374.
[7]张旭.基于微流控液滴的单细胞培养及筛选平台[D].重庆:西南大学,2015.Zhang X.Agarose beads-based long-term single-cell culture and sorting for rapid gowth haematococcus pluvialis microalgae screening[D].Chongqing:Southwest University,2015.
[8]吕晓庆.微流控和单细胞分选技术在生物医学中的应用研究[D].天津:天津医科大学,2015.LüXQ.Research of microfluidic and single cell sorting technology in biomedical applications[D].Tianjin:Tianjin Medical University,2015.
[9]马丛丛.用于单细胞分析的微流控芯片研究及其在肿瘤干细胞分离鉴定方面的应用[D].浙江:浙江大学,2016.Ma CC.The research of microfluidic chip for single cell analysis and its application in isolation and identification of cancer stem cells[D].Zhejiang:Zhejiang University,2016.
[10]王桐.干细胞微流控芯片的设计、制备、检测与应用研究[D].北京:北京工业大学,2013.Wang T.Reasearch on design,fabrication,detection and application of microfluidic biochips for stem cells[D].Beijing:Beijing University of Technology,2013.
[11]Miller KF,Goldberg JM,Collins RL.Covering embryo cultures with mineral oil alters embryo growth by acting as a sink for an embryotoxic substance[J].J Assist Reprod Genet,1994,11(7):342-345.
[12]霍龙,罗奋华,张岩,等.大鼠精原干细胞的微滴培养法[J].中国细胞生物学学报,2014,36(5):671-678.Huo L,Luo FH,Zhang Y,et al.Microdrop culture of rat spermatogonial stem cells[J].Chinese Journal of Cell Biology,2014,36(5):671-678.
[13]蔡志明,石敏,龙云,等.人睾丸圆形精子细胞在体外向长形精子细胞分化[J].中华男科学杂志,2006,12(7):587-589,593.Cai ZM,Shi M,Long Y,et al.In vitro differentiation of human testicular round spermatids to elongating spermatids[J].National Journal of Andrology,2006,12(7):587-589,593.
[14]禹学礼,邓雯,庞有志,等.颗粒细胞和培养微滴大小对牛卵母细胞体外受精后胚胎发育的影响[J].河南农业科学,2005,(9):87-90.Yu XL,Deng W,Pang YZ,et al.Effects of granulosa cell and medium drop size on embryonic development of bovine oocytes following fertilization in vitro[J].Journal of Henan Agricultural Sciences,2005,(9):87-90.
[15]霍龙,张岩,吴应积.微滴培养技术的新应用[J].生物技术通报,2011,(9):59-62.Huo L,Zhang Y,Wu YJ.The new applications of micro-drop culture technique[J].Biotechnology Bulletin,2011,(9):59-62.
[16]Weaver JC,Bliss JG,Powell KT,et al.Rapid clonal growth measurements at the single-cell level:gel microdroplets and flow cytometry[J].Biotechnology(N Y),1991,9(9):873-877.
[17]黄海娟,柴向华,吴克刚.食用油中过氧化物去除方法的研究[C].广州:广东省食品学会第六次会员大会暨学术研讨会论文集,2012.Huang HJ,Chai XH,Wu KG.Study on the methods for removing peroxides in edible oils[C].Proceedings of the Sixth Guangdong Food Institute General Meeting and Symposium,Guangzhou,2012.
[18]罗曌.简析厌氧菌的临床检验[J].世界最新医学信息文摘(连续型电子期刊),2016,16(43):169-170.Luo Z.Clinical laboratory analysis of anaerobes[J].World Latest Medicine Information(Electronic Version),2016,16(43):169-170.
[19]B?rner RA.Isolation and cultivation of anaerobes[J].Adv Biochem Eng Biotechnol,2016,156:35-53.
[20]金艳,张宏,乔建军.两种培养基体外诱导白念珠菌菌丝相形成的比较[J].中华皮肤科杂志,2005,38(8):40-41.Jin Y,Zhang H,Qiao JJ.Induction of hyphal form of Candida albicans in vitro by two culture media[J].Chin J Dermatol,2005,38(8):40-41.
[21]吴幼玲.几种沙氏培养基真菌生长情况比较[J].海峡预防医学杂志,2014,20(4):46-47.Wu YL.Comparison of fungi growth condition in several types of sabouraud mediums[J].Strait J Prev Med,2014,20(4):46-47.
[22]周村建,郝飞,王鲁,等.白念珠菌菌丝的诱导培养[J].第三军医大学学报,2004,26(9):832-833.Zhou CJ,Hao F,Wang L,et al.Hypha formation of Candida albicans by induced culture[J].Acta Academiae Medicinae Militaris Tertiae,2004,26(9):832-833.
[2]蒋析文,朱小亚,高秀洁,等.数字PCR在几种常见人类传染病研究中的应用[J].分子诊断与治疗杂志,2017,9(1):50-54.Jiang XW,Zhu XY,Gao XJ,et al.The applications of digital PCR on the researches of several common human infections[J].J Mol Diagn Ther,2017,9(1):50-54.
[3]赵新,兰青阔,陈锐,等.应用微滴数字PCR技术快速检测食用菌中沙门氏菌[J].食品与生物技术学报,2017,36(3):315-321.Zhao X,Lan QK,Chen R,et al.Rapid detection of Salmonellla spp.in edible fungi by droplet digital PCR[J].Journal of Food Science and Biotechnology,2017,36(3):315-321.
[4]丰蕾,杨帆,李彩霞,等.单细胞分离检验技术在法医学中的应用研究[J].刑事技术,2015,40(5):359-363.Feng L,Yang F,Li CX,et al.Single cell separation testing and its forensic application[J].Forensic Science and Technology,2015,40(5):359-363.
[5]姚金光,解继胜,李俊,等.舌癌单细胞培养建系与癌干细胞相关标志的检测[J].华西口腔医学杂志,2013,31(1):86-90.Yao JG,Xie JS,Li J,et al.Establishment of subseries cell lines from tongue cancer single cell and detection of cancer stem cell markers[J].West China Journal of Stomatology,2013,31(1):86-90.
[6]占爱瑶,由香玲,詹亚光,等.白桦单细胞培养体系的建立[J].植物生理学通讯,2010,46(4):370-374.Zhan AY,You XL,Zhan YG,et al.Establishment of single cell culture of white birch(betula platyphylla suk)[J].Plant Physiology Communications,2010,46(4):370-374.
[7]张旭.基于微流控液滴的单细胞培养及筛选平台[D].重庆:西南大学,2015.Zhang X.Agarose beads-based long-term single-cell culture and sorting for rapid gowth haematococcus pluvialis microalgae screening[D].Chongqing:Southwest University,2015.
[8]吕晓庆.微流控和单细胞分选技术在生物医学中的应用研究[D].天津:天津医科大学,2015.LüXQ.Research of microfluidic and single cell sorting technology in biomedical applications[D].Tianjin:Tianjin Medical University,2015.
[9]马丛丛.用于单细胞分析的微流控芯片研究及其在肿瘤干细胞分离鉴定方面的应用[D].浙江:浙江大学,2016.Ma CC.The research of microfluidic chip for single cell analysis and its application in isolation and identification of cancer stem cells[D].Zhejiang:Zhejiang University,2016.
[10]王桐.干细胞微流控芯片的设计、制备、检测与应用研究[D].北京:北京工业大学,2013.Wang T.Reasearch on design,fabrication,detection and application of microfluidic biochips for stem cells[D].Beijing:Beijing University of Technology,2013.
[11]Miller KF,Goldberg JM,Collins RL.Covering embryo cultures with mineral oil alters embryo growth by acting as a sink for an embryotoxic substance[J].J Assist Reprod Genet,1994,11(7):342-345.
[12]霍龙,罗奋华,张岩,等.大鼠精原干细胞的微滴培养法[J].中国细胞生物学学报,2014,36(5):671-678.Huo L,Luo FH,Zhang Y,et al.Microdrop culture of rat spermatogonial stem cells[J].Chinese Journal of Cell Biology,2014,36(5):671-678.
[13]蔡志明,石敏,龙云,等.人睾丸圆形精子细胞在体外向长形精子细胞分化[J].中华男科学杂志,2006,12(7):587-589,593.Cai ZM,Shi M,Long Y,et al.In vitro differentiation of human testicular round spermatids to elongating spermatids[J].National Journal of Andrology,2006,12(7):587-589,593.
[14]禹学礼,邓雯,庞有志,等.颗粒细胞和培养微滴大小对牛卵母细胞体外受精后胚胎发育的影响[J].河南农业科学,2005,(9):87-90.Yu XL,Deng W,Pang YZ,et al.Effects of granulosa cell and medium drop size on embryonic development of bovine oocytes following fertilization in vitro[J].Journal of Henan Agricultural Sciences,2005,(9):87-90.
[15]霍龙,张岩,吴应积.微滴培养技术的新应用[J].生物技术通报,2011,(9):59-62.Huo L,Zhang Y,Wu YJ.The new applications of micro-drop culture technique[J].Biotechnology Bulletin,2011,(9):59-62.
[16]Weaver JC,Bliss JG,Powell KT,et al.Rapid clonal growth measurements at the single-cell level:gel microdroplets and flow cytometry[J].Biotechnology(N Y),1991,9(9):873-877.
[17]黄海娟,柴向华,吴克刚.食用油中过氧化物去除方法的研究[C].广州:广东省食品学会第六次会员大会暨学术研讨会论文集,2012.Huang HJ,Chai XH,Wu KG.Study on the methods for removing peroxides in edible oils[C].Proceedings of the Sixth Guangdong Food Institute General Meeting and Symposium,Guangzhou,2012.
[18]罗曌.简析厌氧菌的临床检验[J].世界最新医学信息文摘(连续型电子期刊),2016,16(43):169-170.Luo Z.Clinical laboratory analysis of anaerobes[J].World Latest Medicine Information(Electronic Version),2016,16(43):169-170.
[19]B?rner RA.Isolation and cultivation of anaerobes[J].Adv Biochem Eng Biotechnol,2016,156:35-53.
[20]金艳,张宏,乔建军.两种培养基体外诱导白念珠菌菌丝相形成的比较[J].中华皮肤科杂志,2005,38(8):40-41.Jin Y,Zhang H,Qiao JJ.Induction of hyphal form of Candida albicans in vitro by two culture media[J].Chin J Dermatol,2005,38(8):40-41.
[21]吴幼玲.几种沙氏培养基真菌生长情况比较[J].海峡预防医学杂志,2014,20(4):46-47.Wu YL.Comparison of fungi growth condition in several types of sabouraud mediums[J].Strait J Prev Med,2014,20(4):46-47.
[22]周村建,郝飞,王鲁,等.白念珠菌菌丝的诱导培养[J].第三军医大学学报,2004,26(9):832-833.Zhou CJ,Hao F,Wang L,et al.Hypha formation of Candida albicans by induced culture[J].Acta Academiae Medicinae Militaris Tertiae,2004,26(9):832-833.