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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

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Table 2

Polymerase chain reaction (PCR) primers and conditions for preparing DNA probes used in the colony hybridization test

Nucleotide sequence of primers Target PCR conditionsa
Denaturing Annealing Extension Amplicon (bp)
5-GAGAATTCACAGATGGATATTTCAAATTTC-3
5-TCTCCTCGAGCTAGTTGACCTCGTTCAGAAACG-3 espP 94°C, 10s 68°C, 30s 72°C, 90s 2,900
5-GAGAGAATTCTTCCTGTTCTGATTCTTCTG-3
5-TCTCCTCGAGTCAAAACTTATTTCTCGCATCATCATCC-3 katP 94°C, 10s 68°C, 30s 72°C, 60s 2,130
5-TCTCAAGCTTTTAGAAATAGTCTCGCCAGTATTC-3
5-GAGAGAATTCGTCAGGAGGATGTTCAGG-3 eae 94°C, 10s 68°C, 30s 72°C, 90s 880
5-GAGAGGATCCGGTGCAGCAGAAAAAGTTGTA-3
5-TCTCCTCGAGTCATCTCGCCTGATAGTGTTTGG-3 hlyA 94°C, 10s 68°C, 30s 72°C, 60s 1,560

aPCR was done for 30 cycles.

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