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Stenotrophomonas maltophilia  is a common nosocomial pathogen that causes high morbidity and mortality. Because of its inherent extended antibiotic resistance, therapeutic options for  S. maltophilia  are limited, and sulfamethoxazole/trimethoprim (SXT) is the only first-line antimicrobial recommended. However, with the spread of dihydropteroate synthase ( sul1  and  sul2 ) genes, global emergence of SXT resistance has been reported. There is an urgent need to develop a rapid and sensitive but cost-efficient method to monitor the dissemination of  sul  genes. In this study, we developed loop-mediated isothermal amplification (LAMP) assays for  sul1  and  sul2  using real-time turbidity and hydroxy naphthol blue coloration methods. The assays could quickly detect  sul  genes with high sensitivity and specificity. The LAMP detection limit was 0.74 pg/reaction of extracted genomic DNA for  sul1  and 2.6 pg/reaction for  sul2 , which were both 10-fold more sensitive than the corresponding traditional PCR assays. Additionally, the LAMP assays could positively amplify DNA from  sul1 -producing strains, but not from the negative controls. We then used the LAMP assays to investigate the dissemination of  sul  genes among  S. maltophilia  isolates from patients in three hospitals in Beijing, China. Among 450 non-duplicated samples collected during 2012–2014, 56 (12.4%) strains were SXT-resistant. All these SXT-resistant strains were positive for  sul  genes, with 35 (62.5%) carrying  sul1 , 17 (30.4%) carrying  sul2 , and 4 (7.1%) carrying both  sul1  and  sul2 , which indicated that  sul  genes were the predominant resistance mechanism. Of 394 SXT-susceptible strains, 16 were also  sul -positive. To provide epidemiological data for the appropriate choice of antimicrobials for treatment of  sul -positive  S. maltophilia , we further tested the susceptibility to 18 antimicrobials. Among these,  sul -positive strains showed the highest susceptibility to tetracycline derivatives, especially minocycline (MIC 50 /MIC 90 , 0.5/4; susceptibility rate, 95.4%). Ticarcillin-clavulanate and new fluoroquinolones (moxifloxacin and levofloxacin) also showed some  in vitro  activity. Apart from these three kinds of antimicrobials, other agents showed poor activity against  sul -positive strains.

Surveillance of Dihydropteroate Synthase Genes in Stenotrophomonas maltophilia by LAMP: Implications for Infection Control and Initial Therapy