Establishment of a rapid detection method for carbapenem and quinolone resistant nucleic acid colloidal gold test strips and development of a reagent kit
10.3969/j.issn.1000-484X.2024.11.024
- VernacularTitle:碳青霉烯类和喹诺酮类双重耐药核酸胶体金试纸条快速检测方法的建立及试剂盒研制
- Author:
Beizhen PAN
1
;
Jifei YANG
;
Yuefeng WANG
;
Yan LIU
;
Yujiao ZHOU
;
Yuhe MA
;
Liyuan SUN
Author Information
1. 北华大学医学技术学院临床病原学检验教研室,吉林 132013
- Keywords:
Acinetobacter baumannii;
OXA;
par C;
Molecular cloning;
Colloidal gold;
Nucleic acid test strip
- From:
Chinese Journal of Immunology
2024;40(11):2386-2390,2398
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a method for rapid detection of OXA and par C resistance genes of Acinetobacter baumannii(Ab)by double nucleic acid colloidal gold strip and to develop kit.Methods:DNA of Ab was extracted by heating and boiling method.OXA and par C genes sequences of Ab were selected as target gene fragments based on NCBI.Primers were designed and labeled with 6-FAM,digoxin and biotin,respectively.Drug resistance gene detection reagents were developed,and nucleic acid gold test strips were used for rapid and visual detection.Molecular cloning and sequencing techniques were used to clone positive control samples and evaluate specificity,sensitivity and stability of kit.Results:DNA concentration and purity of Ab extracted by boiling method were good.Homology between cloned and sequenced plasmid DNA and gene sequence in GenBank database was 100%,respectively.Speci-ficity of kit was good,with only Ab showing positive results and other bacterial genera showing negative results;DNA concentration of Ab in double nucleic acid colloidal gold test strip decreased to 10-3 ng/μl,a red line still appeared,whose sensitivity was 10 times higher consistent with minimum detection limit of electrophoresis 10-2 ng/μl;test kits were tested at 3rd,6th and 9th months,and showed good stability.Conclusion:Double resistance detection kit established in this study can simultaneously detect OXA and par C resis-tance of Ab,who has advantages of high sensitivity,strong specificity,rapid and simple,and provides a new method for detection of carbapenem and quinolone antibiotic resistance of Ab.
