Publication:
Comparative study on DNA amplification methods for detection of Carbapenem-Resistant Enterobacteriaceae (CRE)

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Issued Date

2021

Resource Type

Article

Language

eng

File Type

application/pdf

ISSN

1252208

DOI

10.35755/jmedassocthai.2021.08.12818

Other identifier(s)

2-s2.0-85113322132

Rights Holder(s)

มหาวิทยาลัยศรีนครินทรวิโรฒ

Bibliographic Citation

Journal of the Medical Association of Thailand. Vol 104, No.8 (2021), p.1293-1300

Suggested Citation

Pakdeethai S., Tantisiriwat W., Areekit S., Bunroddith K., Chansiri K., Santiwatanakul S. Comparative study on DNA amplification methods for detection of Carbapenem-Resistant Enterobacteriaceae (CRE). Journal of the Medical Association of Thailand. Vol 104, No.8 (2021), p.1293-1300. doi:10.35755/jmedassocthai.2021.08.12818 Retrieved from: https://hdl.handle.net/20.500.14740/7994

Title

Comparative study on DNA amplification methods for detection of Carbapenem-Resistant Enterobacteriaceae (CRE)

Author(s)

Pakdeethai S.
 Tantisiriwat W.
 Areekit S.
 Bunroddith K.
 Chansiri K.
 Santiwatanakul S.

Abstract

Background: Carbapenem-resistant Enterobacteriaceae (CRE) is a resistant group of gram-negative bacteria that produces carbapenemase destroying carbapenem molecules causing drug resistances. Among them, the incidence of New Delhi metallo-beta-lactamases (blaNDM-1), a metallocarbapenemases or class B carbapenemases or metallo-β-lactamases (MBL), are increasing worldwide. Objective: To focused on comparative study on DNA amplification methods for detection of CRE in terms of analytical sensitivity and specificity. Materials and Methods: CRE strains (Escherichia coli, Escherichia cloacae, Citrobacter freudii, and Klebsiella pneumonia) were collected from HRH Maha Chakri Sirindhorn Medical Center, Srinakharinwirot University, Nakhon Nayok, Thailand. All specimens were initially screened for CRE isolates by using carbapenem disks inhibition test. The polymerase chain reaction (PCR), loop mediated isothermal amplification (LAMP) and recombinant polymerase amplification (RPA) assays were further employed for identification of drug resistance blaNDM-1 gene among CRE isolates. The analytical sensitivity and specificity of the three amplification assays were compared and analyzed. Results: The analytical sensitivity test of PCR, LAMP and RPA assays revealed that the limit of detection were 0.74 ng/μL, 7.4 pg/μL, and 0.74 ng/μL, respectively. Specificity test demonstrated that all three assays showed no cross hybridization to the other 15 related bacterial isolates. Conclusion: The data pointed out that LAMP assay was 100 times more sensitive than PCR and RPA, which could be suitable for application as early detection test for carbapenem resistant bacteria. Hence, the physician can select the proper antibiotics to treat the CRE infectious bacteria, which can reduce morbidity and mortality rates in patients. In the future, LAMP will be combined with lateral flow dipstick (LFD) to improve the efficacy of the assay as not only the early detection test with high sensitivity and specificity for carbapenem resistant bacteria, but also as the convenient and rapid screening test. © JOURNAL OF THE MEDICAL ASSOCIATION OF THAILAND.

Subject(s)

Beta lactamase
 Carbapenemase
 RNA 16S
 Antibiotic resistance
 Antibiotic sensitivity
 Article
 Bacterial gene
 Bacterial virulence
 Bacterium detection
 Bacterium identification
 Bacterium isolate
 Carbapenem-resistant Enterobacteriaceae
 Comparative study
 Controlled study
 Cross hybridization
 Diagnostic test accuracy study
 Disk diffusion
 DNA extraction
 Drug efficacy
 Drug therapy
 Enzyme synthesis
 Gene amplification
 Klebsiella pneumoniae
 Limit of detection
 Loop mediated isothermal amplification
 Minimum inhibitory concentration
 Mortality
 Mortality rate
 Multidrug resistance
 Multiplex polymerase chain reaction
 Nonhuman
 Nucleotide sequence
 Physician
 Polymerase chain reaction
 Restriction fragment length polymorphism
 Sensitivity and specificity
 Sequence analysis

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URI

https://hdl.handle.net/20.500.14740/7994

Collections

Scopus: Year 1983-2021

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