Development of a serotype-specific DNA microarray for identification of some Shigella and pathogenic Escherichia coli strains.
Shigella and pathogenic Escherichia coli are major causes of human infectious diseases and are responsible for millions of cases of diarrhea worldwide every year. A convenient and rapid method to identify highly pathogenic serotypes of Shigella and E. coli is needed for large-scale epidemiologic study, timely clinical diagnosis, and reliable quarantine of the pathogens. In this study, a DNA microarray targeting O-serotype-specific genes was developed to detect 15 serotypes of Shigella and E. coli, including Shigella sonnei; Shigella flexneri type 2a; Shigella boydii types 7, 9, 13, 16, and 18; Shigella dysenteriae types 4, 8, and 10; and E. coli O55, O111, O114, O128, and O157. The microarray was tested against 186 representative strains of all Shigella and E. coli O serotypes, 38 clinical isolates, and 9 strains of other bacterial species that are commonly present in stool samples and was shown to be specific and reproducible. The detection sensitivity was 50 ng genomic DNA or 10(4) CFU per ml in mock stool specimens. This is the first report of a microarray for serotyping Shigella and pathogenic E. coli. The method has a number of advantages over traditional bacterial culture and antiserum agglutination methods and is promising for applications in basic microbiological research, clinical diagnosis, food safety, and epidemiological surveillance.
Li
Yayue
Y
TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 Hong Da Street, TEDA, Tianjin 300457, China.
Liu
Dan
D
Cao
Boyang
B
Han
Weiqing
W
Liu
Yanqun
Y
Liu
Fenxia
F
Guo
Xi
X
Bastin
David A
DA
Feng
Lu
L
Wang
Lei
L
D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology; D006801:Humans; D009693:Nucleic Acid Hybridization; D019081:O Antigens / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D012680:Sensitivity and Specificity; D012703:Serotyping; D012760:Shigella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
D004269:DNA, Bacterial; D019081:O Antigens