Detection of bacteriophage infection and prophage induction in bacterial cultures by means of electric DNA chips.
Infections of bacterial cultures by bacteriophages are common and serious problems in many biotechnological laboratories and factories. A method for specific, quantitative, and quick detection of phage contamination, based on the use of electric DNA chip is described here. Different phages of Escherichia coli and Bacillus subtilis were analyzed. Phage DNA was isolated from bacterial culture samples and detected by combination of bead-based sandwich hybridization with enzyme-labeled probes and detection of the enzymatic product using silicon chips. The assay resulted in specific signals from all four tested phages without significant background. Although high sensitivity was achieved in 4h assay time, a useful level of sensitivity (10(7)-10(8) phages) is achievable within 25 min. A multiplex DNA chip technique involving a mixture of probes allows for detection of various types of phages in one sample. These analyses confirmed the specificity of the assay.
Gabig-Ciminska
Magdalena
M
Department of Biotechnology, Royal Institute of Technology (KTH), Roslagstullsbacken 21, S-10691 Stockholm, Sweden. magda.gabig@biotech.kth.se
Los
Marcin
M
Holmgren
Anders
A
Albers
Jörg
J
Czyz
Agata
A
Hintsche
Reiner
R
Wegrzyn
Grzegorz
G
Enfors
Sven Olof
SO
D001412:Bacillus subtilis / Q000821:virology; D001419:Bacteria / Q000254:growth & development / Q000821:virology*; D001431:Bacteriological Techniques; D001435:Bacteriophages / Q000302:isolation & purification* / Q000502:physiology; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004926:Escherichia coli / Q000821:virology; D020411:Oligonucleotide Array Sequence Analysis; D013997:Time Factors; D014775:Virus Activation
D015342:DNA Probes; D004279:DNA, Viral