The present paper aims to determine the prevalence of Beijing strains of Mycobacterium tuberculosis isolated from visa applicants in Shanghai Port, to define the probability of drug resistance and to evaluate the underlying risks of dissemination of Mycobacterium tuberculosis and tuberculosis burden for Chinese tuberculosis surveillance system. From January 2009 to May 2013, a total of 193 visa applicants who received medical examinations in Shanghai Port were suspected of infectious tuberculosis by detected abnormalities in the chest radiology. Smear and culture examination from 3 consecutive early morning sputum specimens collected from them were performed. Isolated Mycobacterium tuberculosis complex strains were subjected for genotyping using deletion­targeted multiplex polymerase chain reaction (DTM­PCR). Fifty of them were positive for Mycobacterium tuberculosis complex. Forty isolates were genotyped by DTM­PCR, 23 of the isolates (57.5%) belonged to Beijing genotype. The ratio was significantly lower than that in the local people going abroad (37/41, 90.2%). In all imported cases with Beijing genotype, 5 were from South­East Asia Region and 16 were from Western Pacific Region. Of 23 Beijing strains, 7 (30.4%) were resistant to at least one first­line anti­tuberculosis drug, which was similar to the drug resistance rate (34.1%) in the local people going abroad. No multidrug­resistant Beijing strain was found. The drug resistance rate to pyrazinamide was much higher in visa applicants (16.7%) than that in the local people going abroad (2.4%)．