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Volume 8, Number 1—January 2002
Research

Prevalence and Genetic Profiling of Virulence Determinants of Non-O157 Shiga Toxin-Producing Escherichia coli Isolated from Cattle, Beef, and Humans, Calcutta, India

Asis Khan*, Shinji Yamasaki*†, Toshio Sato†, Thandavarayan Ramamurthy*, Amit Pal*, Simanti Datta*, Nandini Roy Chowdhury*, Suresh Chandra Das‡, Asim Sikdar‡, Teizo Tsukamoto§, Sujit Kumar Bhattacharya*, Yoshifumi Takeda¶, and Gopinath Balakrish Nair*#Comments to Author 
Author affiliations: *National Institute of Cholera and Enteric Diseases, Calcutta, India; †Research Institute, International Medical Center of Japan, Shinjuku-ku, Tokyo, Japan; ‡Indian Veterinary Research Institute, Belgachia, Calcutta, India; §Osaka Prefectural Public Health Institute, Osaka, Japan; ¶National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan; #International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh

Main Article

Table 1

Polymerase chain reaction (PCR) primers and conditions used in this study

Primer no. Nucleotide sequence of primers Target PCR conditionsa
Denaturing Annealing Extension Amplicon (bp) Reference
EVT1/
EVT2 5'-CAACACTGGATGATCTCAG-3'
5'-CCCCCTCAACTGCTAATA-3' stx1 family 94°C, 60s 55°C, 60sb 72°C, 60s 349 20
EVS1/
EVC2 5'-ATCAGTCGTCACTCACTGGT-3'
5'-CTGCTGTCACAGTGACAAA-3' stx2 family 94°C, 60s 55°C, 60sb 72°C, 60s 110 20
hlyA1/
hlyA4 5'-GGTGCAGCAGAAAAAGTTGTAG-3'
5'-TCTCGCCTGATAGTGTTTGGTA-3' EHEC hlyA 94°C, 30s 57°C, 60s 72°C, 90s 1,551 7
wkat-B/
wkat-F 5'-CTTCCTGTTCTGATTCTTCTGG-3'
5'-AACTTATTTCTCGCATCATCC-3' katP 94°C, 30s 56°C, 60s 72°C, 150s 2,125 9
D1/
D13R 5'-CGTCAGGAGGATGTTCAG-3'
5'-CGACTGCACCTGTTCCTGATTA-3' etpD 94°C, 30s 52°C, 60s 72°C, 70s 1,062 21
EAE1/
EAE2 5'-AAACAGGTGAAACTGTTGCC-3'
5'-CTCTGCAGATTAACCTCTGC-3' eae 94°C, 60s 55°C, 90s 72°C, 90s 350 22

a Unless stated, PCR was done for 30 cycles.
b After 35 cycles, final extension step of 10 min at 72°C was performed.

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Page updated: April 18, 2012
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