基于微流控芯片的尿路感染细菌鉴定及抗生素敏感性测试

引用本文: 齐明月, 杜燕, 刘未平, 陈斌, 梁广铁, 徐邦牢, 刘大渔. 基于微流控芯片的尿路感染细菌鉴定及抗生素敏感性测试[J]. 分析化学, 2016, 44(4): 610-616. doi: 10.11895/j.issn.0253-3820.160035 shu

Citation: QI Ming-Yue, DU Yan, LIU Wei-Ping, CHEN Bin, LIANG Guang-Tie, XU Bang-Lao, LIU Da-Yu. Uropathogenic Bacteria Identification and Antibiotic Susceptibility Testing on a Microfluidic Chip[J]. Chinese Journal of Analytical Chemistry, 2016, 44(4): 610-616. doi: 10.11895/j.issn.0253-3820.160035 shu

基于微流控芯片的尿路感染细菌鉴定及抗生素敏感性测试

齐明月 ^1, ,
杜燕 ^1, ,
刘未平 ^1, ,
陈斌 ^1, ,
梁广铁 ^1,2, ,
徐邦牢 ^{1,2,
,} ,
刘大渔 ^1,2,

1.
¹ 广州医科大学附属广州市第一人民医院检验医学研究室, 广州 510180;
2.
² 广东省临床分子医学及分子诊断重点实验室, 广州 510180

通讯作者: 徐邦牢

基金项目:
本文系国家自然科学基金(Nos.81371649,81271730,81501831) (Nos.81371649,81271730,81501831)

广东省自然基金(No.2014A030313677) (No.2014A030313677)

广州市科技计划项目(No.2010U1-E00681)资助 (No.2010U1-E00681)

摘要: 发展了一种微流控芯片纸基细菌分析技术,用于多重细菌鉴定与抗生素敏感性测试。制备了阵列培养池芯片,以滤纸作为衬底固定显色培养基和抗生素。利用PVDF疏水薄膜止流阀,将尿液样品引入芯片并分隔于不同培养池。借助于培养池阵列的空间分辨力,实现多重细菌分析。根据特异性显色结果实现细菌鉴定,通过实时显色强度分析实现细菌定量,依据抑制显色反应的最低抗生素浓度确定抗生素敏感性。以3种泌尿系统感染常见病原菌(大肠杆菌、金黄色葡萄球菌和粪肠球菌)为模拟测试对象进行分析,结果表明,芯片方法可以在18 h内实现对3种细菌的同时鉴定及6种抗生素敏感性测试。对照实验显示,芯片法与传统方法细菌鉴定和抗生素敏感性测试结果一致性分别为94.1%和93.9%。本研究建立的微流控芯片细菌分析方法简便快速,非常适合于医疗资源匮乏条件下的细菌分析。

关键词:

English

Uropathogenic Bacteria Identification and Antibiotic Susceptibility Testing on a Microfluidic Chip

1.
¹ Department of Laboratory Medicine, Guangzhou First People's Hospital, Affiliated Hospital of Guangzhou Medical University, Guangzhou 510180, China;
2.
2

Corresponding author: 徐邦牢

Received Date: 14 January 2016
Fund Project:
本文系国家自然科学基金(Nos.81371649,81271730,81501831)

广东省自然基金(No.2014A030313677)

广州市科技计划项目(No.2010U1-E00681)资助

Abstract: A microfluidic chip assay was described for bacteria identification and antibiotic susceptibility test(AST). Filter paper pads were embedded in arrayed microchambers, with immobilized antibiotics and chromogenic medium. By taking advantage of the polyvinylidene fluoride(PVDF) membrane based valves, urine sample introduced was distributed in individual chambers without cross contamination. The simultaneous analysis of multiple bacteria was achieved by using the culture chamber array design. The identification of a bacterium was based on its specific colorimetric result. The density of a bacterium was determined by real-time monitoring color intensity in the chamber, and its susceptibility to an antibiotic was determined relying on the lowest antibiotic concentration that inhibited the colorimetric reaction. A set of three common uropathogenic bacteria were selected as models to test the microfluidic approach. Our results showed that the developed microfluidic assay was able to complete bacteria identification and the six-antibiotic AST in 18 h. In comparison with the conventional method, the microchip method showed a coincidence of 94.1% and 93.9% with regard to bacteria identification and AST, respectively. The developed microfluidic approach is simple and rapid, thus hold the potential to serve as a powerful tool for bacterial analysis under conditions of poor medical resource.

Key words:

1. [1]
  1 Honrado C, Dong T. Sensors(Basel), 2014, 14(8):13851-138621 Honrado C, Dong T. Sensors(Basel), 2014, 14(8):13851-13862
2. [2]
  2 Mach K E, Wong P K, Liao J C. Trends Pharmacol. Sci., 2011, 32(6):330-3362 Mach K E, Wong P K, Liao J C. Trends Pharmacol. Sci., 2011, 32(6):330-336
3. [3]
  3 Graham J C, Galloway A. J. Clin. Pathol., 2001, 54(12):911-9193 Graham J C, Galloway A. J. Clin. Pathol., 2001, 54(12):911-919
4. [4]
  4 WANG Hong-Ru, BAO Xiao-Rong, YUAN Yi-Qun. Chinese Journal of Nosocomiology, 2014, 24(08):1859-1861 王宏儒, 鲍晓荣, 袁轶群.中华医院感染学杂志, 2014, 24(08):1859-18614 WANG Hong-Ru, BAO Xiao-Rong, YUAN Yi-Qun. Chinese Journal of Nosocomiology, 2014, 24(08):1859-1861 王宏儒, 鲍晓荣, 袁轶群.中华医院感染学杂志, 2014, 24(08):1859-1861
5. [5]
  5 Luo X, Shen K, Luo C, Ji H, Ouyang Q, Chen Y. Biomed. Microdevices, 2010, 12(3):499-5035 Luo X, Shen K, Luo C, Ji H, Ouyang Q, Chen Y. Biomed. Microdevices, 2010, 12(3):499-503
6. [6]
  6 Meister L, Morley E J, Scheer D, Sinert R. Acad. Emerg. Med., 2013, 20(7):631-6456 Meister L, Morley E J, Scheer D, Sinert R. Acad. Emerg. Med., 2013, 20(7):631-645
7. [7]
  7 Matos Pires N M, Dong T. Sensors(Basel), 2013, 13(12):15898-159117 Matos Pires N M, Dong T. Sensors(Basel), 2013, 13(12):15898-15911
8. [8]
  8 Devillé W L, Yzermans J C, van Duijn N P, Bezemer P D, van der Windt D A, Bouter L M. BMC. Urol., 2004, 4:48 Devillé W L, Yzermans J C, van Duijn N P, Bezemer P D, van der Windt D A, Bouter L M. BMC. Urol., 2004, 4:4
9. [9]
  9 Pieretti B, Brunati P, Pini B, Colzani C, Congedo P, Rocchi M, Terramocci R. J. Clin. Microbiol., 2010, 48(11):3990-39969 Pieretti B, Brunati P, Pini B, Colzani C, Congedo P, Rocchi M, Terramocci R. J. Clin. Microbiol., 2010, 48(11):3990-3996
10. [10]
  10 Cornut P L, Boisset S, Romanet J P, Maurin M, Carricajo A, Benito Y, Vandenesch F, Chiquet C. Surv. Ophthalmol., 2014, 59(3):286-30310 Cornut P L, Boisset S, Romanet J P, Maurin M, Carricajo A, Benito Y, Vandenesch F, Chiquet C. Surv. Ophthalmol., 2014, 59(3):286-303
11. [11]
  11 Nakanishi T. Rinsho Byori., 2015, 63(4):465-47111 Nakanishi T. Rinsho Byori., 2015, 63(4):465-471
12. [12]
  12 Fournier P E, Drancourt M, Colson P, Rolain J M, La Scola B, Raoult D. Nat. Rev. Microbiol., 2013, 11(8):574-58512 Fournier P E, Drancourt M, Colson P, Rolain J M, La Scola B, Raoult D. Nat. Rev. Microbiol., 2013, 11(8):574-585
13. [13]
  13 Dong T, Zhao X. Anal. Chem., 2015, 87(4):2410-241813 Dong T, Zhao X. Anal. Chem., 2015, 87(4):2410-2418
14. [14]
  14 Feng S, Dong T, Yang Z. IEEE. Eng. Med. Biol. Soc. Conf. Proc., 2014:3114-311714 Feng S, Dong T, Yang Z. IEEE. Eng. Med. Biol. Soc. Conf. Proc., 2014:3114-3117
15. [15]
  15 Lee W, Kwon D, Choi W, Jung G Y, Jeon S. Sci. Rep., 2015, 5:771715 Lee W, Kwon D, Choi W, Jung G Y, Jeon S. Sci. Rep., 2015, 5:7717
16. [16]
  16 He J, Mu X, Guo Z, Hao H, Zhang C, Zhao Z, Wang Q. Eur. J. Clin. Microbiol. Infect. Dis., 2014, 33(12):2223-223016 He J, Mu X, Guo Z, Hao H, Zhang C, Zhao Z, Wang Q. Eur. J. Clin. Microbiol. Infect. Dis., 2014, 33(12):2223-2230
17. [17]
  17 Kalashnikov M, Campbell J, Lee J C, Sharon A, Sauer-Budge A F. J. Vis. Exp., 2014,(83):e5082817 Kalashnikov M, Campbell J, Lee J C, Sharon A, Sauer-Budge A F. J. Vis. Exp., 2014,(83):e50828
18. [18]
  18 Tokel O, Yildiz U H, Inci F, Durmus N G, Ekiz O O, Turker B, Cetin C, Rao S, Sridhar K, Natarajan N, Shafiee H, Dana A, Demirci U. Sci. Rep., 2015, 5:915218 Tokel O, Yildiz U H, Inci F, Durmus N G, Ekiz O O, Turker B, Cetin C, Rao S, Sridhar K, Natarajan N, Shafiee H, Dana A, Demirci U. Sci. Rep., 2015, 5:9152
19. [19]
  19 Cira N J, Ho J Y, Dueck M E, Weibel D B. Lab Chip, 2012, 12(6):1052-105919 Cira N J, Ho J Y, Dueck M E, Weibel D B. Lab Chip, 2012, 12(6):1052-1059
20. [20]
  20 Funes-Huacca M, Wu A, Szepesvari E, Rajendran P, Kwan-Wong N, Razgulin A, Shen Y, Kagira J, Campbell R, Derda R. Lab Chip, 2012, 12(21):4269-427820 Funes-Huacca M, Wu A, Szepesvari E, Rajendran P, Kwan-Wong N, Razgulin A, Shen Y, Kagira J, Campbell R, Derda R. Lab Chip, 2012, 12(21):4269-4278
21. [21]
  21 Sun P, Liu Y, Sha J, Zhang Z, Tu Q, Chen P, Wang J. Biosens. Bioelectron., 2011, 26(5):1993-199921 Sun P, Liu Y, Sha J, Zhang Z, Tu Q, Chen P, Wang J. Biosens. Bioelectron., 2011, 26(5):1993-1999
22. [22]
  22 Sakamoto C, Yamaguchi N, Nasu M. Appl. Environ. Microbiol., 2005, 71(2):1117-112122 Sakamoto C, Yamaguchi N, Nasu M. Appl. Environ. Microbiol., 2005, 71(2):1117-1121
23. [23]
  23 WEN Xiao-Xia, XU Bang-Lao, WANG Wei-Xin, LIANG Guang-Tie, CHEN Bin, YANG Yin-Mei, LIU Da-Yu. Chinese J. Anal. Chem., 2014, 42(6):791-798 文小霞, 徐邦牢, 王伟鑫, 梁广铁, 陈斌, 杨银梅, 刘大渔. 分析化学, 2014, 42(6):791-79823 WEN Xiao-Xia, XU Bang-Lao, WANG Wei-Xin, LIANG Guang-Tie, CHEN Bin, YANG Yin-Mei, LIU Da-Yu. Chinese J. Anal. Chem., 2014, 42(6):791-798 文小霞, 徐邦牢, 王伟鑫, 梁广铁, 陈斌, 杨银梅, 刘大渔. 分析化学, 2014, 42(6):791-798
24. [24]
  24 Cate D M, Adkins J A, Mettakoonpitak J, Henry C S. Anal. Chem., 2015, 87(1):19-4124 Cate D M, Adkins J A, Mettakoonpitak J, Henry C S. Anal. Chem., 2015, 87(1):19-41
25. [25]
  25 Stedtfeld R D, Tourlousse D M, Seyrig G, Stedtfeld T M, Kronlein M, Price S, Ahmad F, Gulari E, Tiedje J M, Hashsham S A. Lab Chip., 2012, 12(8):1454-146225 Stedtfeld R D, Tourlousse D M, Seyrig G, Stedtfeld T M, Kronlein M, Price S, Ahmad F, Gulari E, Tiedje J M, Hashsham S A. Lab Chip., 2012, 12(8):1454-1462
26. [26]
  26 Jokerst J C, Adkins J A, Bisha B, Mentele M M, Goodridge L D, Henry C S. Anal. Chem., 2012, 84(6):2900-290726 Jokerst J C, Adkins J A, Bisha B, Mentele M M, Goodridge L D, Henry C S. Anal. Chem., 2012, 84(6):2900-2907
27. [27]
  27 Clinicaland Laboratory Standards Institute, Performance Standards for Antimicrobial Susceptibility Testing. Twenty-Third Informational Supplement. www.clsi.org, 201327 Clinicaland Laboratory Standards Institute, Performance Standards for Antimicrobial Susceptibility Testing. Twenty-Third Informational Supplement. www.clsi.org, 2013

扫一扫看文章

计量

PDF下载量: 1
文章访问数: 855
HTML全文浏览量: 53

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于微流控芯片的尿路感染细菌鉴定及抗生素敏感性测试

通讯作者: 徐邦牢

English

Uropathogenic Bacteria Identification and Antibiotic Susceptibility Testing on a Microfluidic Chip

Corresponding author: 徐邦牢

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于微流控芯片的尿路感染细菌鉴定及抗生素敏感性测试

通讯作者: 徐邦牢

English

Uropathogenic Bacteria Identification and Antibiotic Susceptibility Testing on a Microfluidic Chip

Corresponding author: 徐邦牢

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs