Filter-Based Assay for Escherichia coli in Aqueous Samples Using Bacteriophage-Based Amplification

被引:61
作者
Derda, Ratmir [1 ,2 ,3 ,4 ]
Lockett, Matthew R. [1 ]
Tang, Sindy K. Y. [1 ,2 ]
Fuller, Renee C. [1 ]
Maxwell, E. Jane [1 ]
Breiten, Benjamin [1 ]
Cuddemi, Christine A. [1 ]
Ozdogan, Aysegul [1 ]
Whitesides, George M. [1 ,2 ]
机构
[1] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA
[2] Harvard Univ, Wyss Inst Biol Inspired Engn, Cambridge, MA 02138 USA
[3] Univ Alberta, Dept Chem, Edmonton, AB T6G 2G2, Canada
[4] Univ Alberta, Alberta Glyc Ctr, Edmonton, AB T6G 2G2, Canada
基金
比尔及梅琳达.盖茨基金会; 美国国家科学基金会;
关键词
BETA-GALACTOSIDASE; PATHOGEN DETECTION; ALPHA-COMPLEMENTATION; PHAGE; CONSTRUCTION; O157-H7; IDENTIFICATION; TUBERCULOSIS; SENSITIVITY; LIBRARIES;
D O I
10.1021/ac400961b
中图分类号
O65 [分析化学];
学科分类号
070302 [分析化学];
摘要
This paper describes a method to detect the presence of bacteria in aqueous samples, based on the capture of bacteria on a syringe filter, and the infection of targeted bacterial species with a bacteriophage (phage). The use of phage as a reagent provides two opportunities for signal amplification: (i) the replication of phage inside a live bacterial host and (ii) the delivery and expression of the complementing gene that turns on enzymatic activity and produces a colored or fluorescent product. Here we demonstrate a phage-based amplification scheme with an M13KE phage that delivers a small peptide motif to an F+, alpha-complementing strain of Escherichia coli K12, which expresses the omega-domain of beta-galactosidase (beta-gal). The result of this complementation-an active form of beta-gal-was detected colorimetrically, and the high level of expression of the omega-domain of beta-gal in the model K12 strains allowed us to detect, on average, five colony-forming units (CFUs) of this strain in 1 L of water with an overnight culture-based assay. We also detected SO CFUs of the model K12 strain in 1 L of water (or 10 mL of orange juice, or 10 mL of skim milk) in less than 4 h with a solution-based assay with visual readout. The solution-based assay does not require specialized equipment or access to a laboratory, and is more rapid than existing tests that are suitable for use at the point of access. This method could potentially be extended to detect many different bacteria with bacteriophages that deliver genes encoding a full-length enzyme that is not natively expressed in the target bacteria.
引用
收藏
页码:7213 / 7220
页数:8
相关论文
共 45 条
[1]
[Anonymous], 2002, EPA821R02020
[2]
[Anonymous], 2011, PRIV DRINK WAT WELLS
[3]
[Anonymous], 2008, FACT DRINK WAT GROUN
[4]
[Anonymous], 2012, NCBI GEN RES
[5]
[Anonymous], 2001, Phage Display: A Laboratory Manual
[6]
Evaluation of a semi-automated reporter phage assay for susceptibility testing Myobacterium tuberculosis isolates in South Africa [J].
Banaiee, Niaz ;
January, Vanessa ;
Barthus, Charmaine ;
Lambrick, Maureen ;
RoDiti, Denise ;
Behr, Marcel A. ;
Jacobs, William R., Jr. ;
Steyn, Lafras M. .
TUBERCULOSIS, 2008, 88 (01) :64-68
[7]
Materials science - Food pathogen detection [J].
Batt, Carl A. .
SCIENCE, 2007, 316 (5831) :1579-1580
[8]
Calendar R., 2006, BACTERIOPHAGES
[9]
Diversity of Phage-Displayed Libraries of Peptides during Panning and Amplification [J].
Derda, Ratmir ;
Tang, Sindy K. Y. ;
Li, S. Cory ;
Ng, Simon ;
Matochko, Wadim ;
Jafari, Mohammad R. .
MOLECULES, 2011, 16 (02) :1776-1803
[10]
Uniform Amplification of Phage with Different Growth Characteristics in Individual Compartments Consisting of Monodisperse Droplets [J].
Derda, Ratmir ;
Tang, Sindy K. Y. ;
Whitesides, George M. .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2010, 49 (31) :5301-5304