Development of a One-Step Multiplex PCR Assay for Differential Detection of Four species (Enterobacter cloacae, Enterobacter hormaechei, Enterobacter roggenkampii, and Enterobacter kobei) Belonging to Enterobacter cloacae Complex With Clinical Significance

Ji, Yang1; Wang, Peihong2,3,4; Xu, Tingting1; Zhou, Yanzi5; Chen, Rongchang1; Zhu, Huaiqiu2,3,4; Zhou, Kai1

2021-05-18

10.3389/fcimb.2021.677089

Frontiers in Cellular and Infection Microbiology   影响因子和分区

2235-2988

Enterobacter cloacae complex (ECC) is composed of multiple species and the taxonomic status is consecutively updated. In last decades ECC is frequently associated with multidrug resistance and become an important nosocomial pathogen. Currently, rapid and accurate identification of ECC to the species level remains a technical challenge, thus impedes our understanding of the population at the species level. Here, we aimed to develop a simple, reliable, and economical method to distinguish four epidemiologically prevalent species of ECC with clinical significance, i.e., E. cloacae, E. hormaechei, E. roggenkampii, and E. kobei. A total of 977 ECC genomes were retrieved from the GenBank, and unique gene for each species was obtained by core-genome comparisons. Four pairs of species-specific primers were designed based on the unique genes. A total of 231 ECC clinical strains were typed both by hsp60 typing and by species-specific PCRs. The specificity and sensitivity of the four species-specific PCRs ranged between 96.56% and 100% and between 76.47% and 100%, respectively. The PCR for E. cloacae showed the highest specificity and sensitivity. A one-step multiplex PCR was subsequently established by combining the species-specific primers. Additional 53 hsp60-typed ECC and 20 non-ECC isolates belonging to six species obtained from samples of patients, sewage water and feces of feeding animals were tested by the multiplex PCR. The identification results of both techniques were concordant. The multiplex PCR established in this study provides an accurate, expeditious, and cost-effective way for routine diagnosis and molecular surveillance of ECC strains at species level.

multiplex PCR Enterobacter cloacae complex Enterobacter cloacae Enterobacter hormaechei Enterobacter roggenkampii and Enterobacter kobei

SCI

英语

第一 ; 通讯

National Key Research and Development Program of China[2017YFC1200200] ; Major Infectious Diseases Such as AIDS and Viral Hepatitis Prevention and Control Technology Major Projects[2018ZX10712-001] ; National Natural Science Foundation of China[81702045,81902030] ; Shenzhen Basic Research projects[JCYJ20190807144409307]

Immunology ; Microbiology

Immunology ; Microbiology

WOS:000656943800001

FRONTIERS MEDIA SA

Web of Science

1.Southern Univ Sci & Technol, Shenzhen Peoples Hosp, Affiliated Hosp 1, Shenzhen Inst Resp Dis,Clin Med Coll 2,Jinan Univ, Shenzhen, Peoples R China
2.Peking Univ, Coll Engn, State Key Lab Turbulence & Complex Syst, Beijing, Peoples R China
3.Peking Univ, Coll Engn, Dept Biomed Engn, Beijing, Peoples R China
4.Peking Univ, Ctr Quantitat Biol, Beijing, Peoples R China
5.Zhejiang Univ, Coll Med, Affiliated Hosp 1, State Key Lab Diag & Treatment Infect Dis,Collabo, Hangzhou, Peoples R China

Ji, Yang,Wang, Peihong,Xu, Tingting,et al. Development of a One-Step Multiplex PCR Assay for Differential Detection of Four species (Enterobacter cloacae, Enterobacter hormaechei, Enterobacter roggenkampii, and Enterobacter kobei) Belonging to Enterobacter cloacae Complex With Clinical Significance[J]. Frontiers in Cellular and Infection Microbiology,2021,11.

Ji, Yang.,Wang, Peihong.,Xu, Tingting.,Zhou, Yanzi.,Chen, Rongchang.,...&Zhou, Kai.(2021).Development of a One-Step Multiplex PCR Assay for Differential Detection of Four species (Enterobacter cloacae, Enterobacter hormaechei, Enterobacter roggenkampii, and Enterobacter kobei) Belonging to Enterobacter cloacae Complex With Clinical Significance.Frontiers in Cellular and Infection Microbiology,11.

Ji, Yang,et al."Development of a One-Step Multiplex PCR Assay for Differential Detection of Four species (Enterobacter cloacae, Enterobacter hormaechei, Enterobacter roggenkampii, and Enterobacter kobei) Belonging to Enterobacter cloacae Complex With Clinical Significance".Frontiers in Cellular and Infection Microbiology 11(2021).