Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii

Background:Acinetobacter baumannii(A.baumannii)is one of the pivotal pathogens responsible for nosocomial infections,especially in patients with low immune response,and infection with carbapenem-resistant A.baumannii has been increasing in recent years.Rapid and accurate detection of carbapenem-resistance genes in A.baumannii could be of immense help to clinical staff.Methods:In this study,a 15-μL reaction system for recombinase polymerase amplification(RPA)was developed and tested.We collected 30 clinical isolates of A.baumannii from the Burn Institute of Southwest Hospital of Third Military Medical University(Army Medical University)for 6 months and tested antibiotic susceptibility using the VITEK 2 system.A.baumannii was detected based on the blaOXA-51 gene by PCR,qPCR and 15μL-RPA,respectively.Sensitivity and specificity were evaluated.In addition,PCR and 15μL-RPA data for detecting the carbapenem-resistance gene blaOXA-23 were comparatively assessed.Results:The detection limit of the blaOXA-51 gene by 15μL RPA was 2.86 CFU/ml,with sensitivity comparable to PCR and qPCR.No positive amplification signals were detected in non-Acinetobacter isolates,indicating high specificity.However,only 18 minutes were needed for the 15μL RPA assay.Furthermore,an antibiotic susceptibility test showed that up to 90%of A.baumannii strains were resistant to meropenem and imipenem;15μL RPA data for detecting blaOXA-23 showed that only 10%(n=3)of A.baumannii isolates did not show positive amplification signals,and the other 90%of(n=27)isolates were positive,corroborating PCR results.Conclusion:We demonstrated that the new 15μL RPA assay for detecting blaOXA-23 in A.baumannii is faster and simpler than qPCR and PCR.It is a promising alternative molecular diagnostic tool for rapid and effective detection of A.baumannii and drug-resistance genes in the field and point-ofcare testing.

Bioprinting of skin constructs for wound healing

Extensive burns and full-thickness skin wounds are difficult to repair. Autologous split-thickness skin graft (ASSG) is still used as the gold standard in the clinic. However, the shortage of donor skin tissues is a serious problem. A potential solution to this problem is to fabricate skin constructs using biomaterial scaffolds with or without cells. Bioprinting is being applied to address the need for skin tissues suitable for transplantation, and can lead to the development of skin equivalents for wound healing therapy. Here, we summarize strategies of bioprinting and review current advances of bioprinting of skin constructs. There will be challenges on the way of 3D bioprinting for skin regeneration, but we still believe bioprinting will be potential skills for wounds healing in the foreseeable future.