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Microorganisms
Volume 13
Issue 11
10.3390/microorganisms13112590
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Article

Rapid 65-min SYBR-Green PCR Assay for Carbapenem Resistant Klebsiella and Acinetobacter Detection

by
Sebnem Emine Bukavaz
1,*,
Kultural Gungor
2,
Hakan Kunduracılar
1 and
Zerrin Yulugkural
3
1
Health Services Vocational College, Trakya University, 22030 Edirne, Turkey
2
Department of Infectious Diseases and Clinical Microbiology, Kırklareli University, 39100 Kırklareli, Turkey
3
Department of Infectious Disease and Clinical Microbiology, Trakya University, 22030 Edirne, Turkey
*
Author to whom correspondence should be addressed.
Microorganisms 2025, 13(11), 2590; https://doi.org/10.3390/microorganisms13112590 (registering DOI)
Submission received: 21 October 2025 / Revised: 11 November 2025 / Accepted: 12 November 2025 / Published: 13 November 2025
(This article belongs to the Special Issue Healthcare-Associated Infections and Antimicrobial Therapy—2nd Edition)
Versions Notes

Abstract

This study developed a rapid and reliable SYBR-Green semiplex PCR assay for simulta-neous detection of major carbapenem resistance genes in Klebsiella pneumoniae and Acinetobacter baumannii. Two primer sets were used: one to detect blaKPC, blaNDM-1, and blaOXA-48 genes in K. pneumoniae and blaOXA-23 in A. baumannii, and another to amplify conserved 16S rRNA gene regions as internal controls. The intra- and inter-assay coeffi-cient of variation ranged from 0.03% to 3.8%. Standard curves exhibited excellent linearity across six logarithmic scales (101–106 DNA copies/µL), with detection limits of 10–102 DNA copies/mL. Melting temperatures (Tm) were: 88.85 °C (KPIC), 90.65 °C (NDM-1), 89.45 °C (KPC), 84.23 °C (OXA-48), 87.81 °C (OXA-23), and 80.67 °C (ABIC). The SYBR-Green Semiplex PCR assay offers a rapid (65 min turnaround), cost-effective, and sensitive method for early detection of carbapenem-resistant pathogens, enabling timely targeted therapy and improved infection control by potentially reducing empirical antibiotic use before culture confirmation.
Keywords: SYBR-Green PCR; Klebsiella pneumoniae; Acinetobacter baumannii SYBR-Green PCR; Klebsiella pneumoniae; Acinetobacter baumannii

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MDPI and ACS Style

Bukavaz, S.E.; Gungor, K.; Kunduracılar, H.; Yulugkural, Z. Rapid 65-min SYBR-Green PCR Assay for Carbapenem Resistant Klebsiella and Acinetobacter Detection. Microorganisms 2025, 13, 2590. https://doi.org/10.3390/microorganisms13112590

AMA Style

Bukavaz SE, Gungor K, Kunduracılar H, Yulugkural Z. Rapid 65-min SYBR-Green PCR Assay for Carbapenem Resistant Klebsiella and Acinetobacter Detection. Microorganisms. 2025; 13(11):2590. https://doi.org/10.3390/microorganisms13112590

Chicago/Turabian Style

Bukavaz, Sebnem Emine, Kultural Gungor, Hakan Kunduracılar, and Zerrin Yulugkural. 2025. "Rapid 65-min SYBR-Green PCR Assay for Carbapenem Resistant Klebsiella and Acinetobacter Detection" Microorganisms 13, no. 11: 2590. https://doi.org/10.3390/microorganisms13112590

APA Style

Bukavaz, S. E., Gungor, K., Kunduracılar, H., & Yulugkural, Z. (2025). Rapid 65-min SYBR-Green PCR Assay for Carbapenem Resistant Klebsiella and Acinetobacter Detection. Microorganisms, 13(11), 2590. https://doi.org/10.3390/microorganisms13112590

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