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We first reported the vanM vancomycin resistance gene in enterococci in Shanghai, China in 2006 and later found it to be the predominant van gene in vancomycin‐resistant enterococci (VRE). In this study, we successively collected 1292 Enterococcus faecium and Enterococcus faecalis strains from in‐ and outpatients at Huashan Hospital, Fudan University and found that nearly all of the isolates (1290/1292) were vancomycin‐sensitive determined by the VITEK 2 system. However, using a modified macromethod‐based disk diffusion test, 10 E. faecium isolates that were previously determined to be vancomycin‐sensitive by the VITEK 2 system were found to have colonies in the vancomycin disk inhibition zone. Pulse‐field gel electrophoresis results showed that each randomly selected colony in the inhibition zone belonged to the same clone as the original strain. All 10 isolates were later found to be vanM‐positive. The disk diffusion‐based method may aid in the detection of vanM‐positive E. faecium with low vancomycin minimum inhibitory concentrations and prevent missing the detection of vancomycin sensitivity‐variable enterococci. [ABSTRACT FROM AUTHOR]

Background: Human rhinoviruses (HRVs) are a highly prevalent cause of acute respiratory infection in children. They are classified into at least three species, HRV-A, HRV-B and HRV-C, which are characterized by sequencing the 59 untranslated region (UTR) or the VP4/VP2 region of the genome. Given the increased interest for novel HRV strain identification and their worldwide distribution, we have carried out clinical and molecular diagnosis of HRV strains in a 2-year study of children with acute respiratory infection visiting one district hospital in Shanghai. Methodology/Findings: We cloned and sequenced a 924-nt fragment that covered part of the 59UTR and the VP4/VP2 capsid genes. Sixty-four HRV-infected outpatients were diagnosed amongst 827 children with acute low respiratory tract infection. Two samples were co-infected with HRV-A and HRV-B or HRV-C. By comparative analysis of the VP4/VP2 sequences of the 66 HRVs, we showed a high diversity of strains in HRV-A and HRV-B species, and a prevalence of 51.5% of strains that belonged to the recently identified HRV-C species. When analyzing a fragment of the 59 UTR, we characterized at least two subspecies of HRV-C: HRV-Cc, which clustered differently from HRV-A and HRV-B, and HRV-Ca, which resulted from previous recombination in this region with sequences related to HRV-A. The full-length sequence of one strain of each HRVCa and HRV-Cc subspecies was obtained for comparative analysis. We confirmed the close relationship of their structural proteins but showed apparent additional recombination events in the 2A gene and 39UTR of the HRV-Ca strain. Double or triple infections with HRV-C and respiratory syncytial virus and/or bocavirus were diagnosed in 33.3% of the HRV-infected patients, but no correlation with severity of clinical outcome was observed. Conclusion: Our study showed a high diversity of HRV strains that cause bronchitis and pneumonia in children. A predominance of HRV-C over HRV-A and HRV-B was observed, and two subspecies of HRV-C were identified, the diversity of which seemed to be related to recombination with former HRV-A strains. None of the HRV-C strains appeared to have a higher clinical impact than HRV-A or HRV-B on respiratory compromise. [ABSTRACT FROM AUTHOR]