Rapid Visual Detection of Methicillin-Resistant in Human Clinical Samples Via Closed LAMP Assay Targeting and Genes

Overview

Journal Microorganisms

Specialty Microbiology

Date 2024 Jan 23

PMID 38257983

Authors

Noora S A Abusheraida

Asraa A H AlBaker

Asmaa S A Aljabri

Hana A Abdelrahman

Hassan Al-Mana

Godwin J Wilson

Khalid A Anan

Nahla O Eltai

Affiliations

Soon will be listed here.

Abstract

The emergence of antimicrobial resistance (AMR), particularly methicillin-resistant (MRSA), poses a significant global health threat as these bacteria increasingly become resistant to the most available therapeutic options. Thus, developing an efficient approach to rapidly screen MRSA directly from clinical specimens has become vital. In this study, we establish a closed-tube loop-mediated isothermal amplification (LAMP) method incorporating hydroxy-naphthol blue (HNB) colorimetric dye assay to directly detect MRSA from clinical samples based on the presence of and genes. In total, 125 preidentified isolates and 93 clinical samples containing were sourced from the microbiology laboratory at Hamad General Hospital (HGH). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were computed based on conventional PCR. The assay demonstrated 100% specificity, 91.23% sensitivity, 0.90 Cohen Kappa (CK), 100% PPV, and 87.8% NPV for the clinical samples, while clinical isolates exhibited 100% specificity, 97% sensitivity, 0.926 CK, 100% PPV, and 88.89% NPV. Compared to cefoxitin disk diffusion, LAMP provided 100% specificity and sensitivity, 1.00 CK, and 100% for PPV and NPV. The study revealed that the closed-tube LAMP incorporating (HNB) dye is a rapid technique with a turnaround time of less than 1 h and high specificity and sensitivity.

References

Nii-Trebi N . Emerging and Neglected Infectious Diseases: Insights, Advances, and Challenges. Biomed Res Int. 2017; 2017:5245021. PMC: 5327784. DOI: 10.1155/2017/5245021. View

Lee A, de Lencastre H, Garau J, Kluytmans J, Malhotra-Kumar S, Peschel A . Methicillin-resistant Staphylococcus aureus. Nat Rev Dis Primers. 2018; 4:18033. DOI: 10.1038/nrdp.2018.33. View

Alipour F, Ahmadi M, Javadi S . Evaluation of different methods to detect methicillin resistance in Staphylococcus aureus (MRSA). J Infect Public Health. 2014; 7(3):186-91. DOI: 10.1016/j.jiph.2014.01.007. View

Zaczek-Moczydlowska M, Mohamed-Smith L, Toldra A, Hooper C, Campas M, Furones M . A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the . Int J Mol Sci. 2020; 21(18). PMC: 7555478. DOI: 10.3390/ijms21186605. View

Zhao L, Huang X, Zhang T, Zhang X, Jiang M, Lu H . A point-of-care test device for MRSA rapid detection. J Pharm Biomed Anal. 2021; 209:114464. DOI: 10.1016/j.jpba.2021.114464. View

Diego J, Fernandez-Soto P, Dominguez-Gil M, Belhassen-Garcia M, Munoz Bellido J, Muro A . A imple, ffordable, apid, tabilized, lorimetric, ersatile RT-LAMP Assay to Detect SARS-CoV-2. Diagnostics (Basel). 2021; 11(3). PMC: 8000859. DOI: 10.3390/diagnostics11030438. View

Trevethan R . Sensitivity, Specificity, and Predictive Values: Foundations, Pliabilities, and Pitfalls in Research and Practice. Front Public Health. 2017; 5:307. PMC: 5701930. DOI: 10.3389/fpubh.2017.00307. View

Durand G, Raoult D, Dubourg G . Antibiotic discovery: history, methods and perspectives. Int J Antimicrob Agents. 2018; 53(4):371-382. DOI: 10.1016/j.ijantimicag.2018.11.010. View

Peacock S, Paterson G . Mechanisms of Methicillin Resistance in Staphylococcus aureus. Annu Rev Biochem. 2015; 84:577-601. DOI: 10.1146/annurev-biochem-060614-034516. View

10.

Bhatt R, Shrestha C, Risal P . Factors Affecting Turnaround Time in the Clinical Laboratory of the Kathmandu University Hospital, Nepal. EJIFCC. 2019; 30(1):14-24. PMC: 6416806. View

11.

Laupland K . Incidence of bloodstream infection: a review of population-based studies. Clin Microbiol Infect. 2013; 19(6):492-500. DOI: 10.1111/1469-0691.12144. View

12.

Al Mana H, Sundararaju S, Tsui C, Perez-Lopez A, Yassine H, Thani A . Whole-Genome Sequencing for Molecular Characterization of Carbapenem-Resistant Enterobacteriaceae Causing Lower Urinary Tract Infection among Pediatric Patients. Antibiotics (Basel). 2021; 10(8). PMC: 8388976. DOI: 10.3390/antibiotics10080972. View

13.

Otto M . MRSA virulence and spread. Cell Microbiol. 2012; 14(10):1513-21. PMC: 3443268. DOI: 10.1111/j.1462-5822.2012.01832.x. View

14.

El-Shishtawy R, Mohamed S, Asiri A, Gomaa A, Ibrahim I, Al-Talhi H . Solid fermentation of wheat bran for hydrolytic enzymes production and saccharification content by a local isolate Bacillus megatherium. BMC Biotechnol. 2014; 14:29. PMC: 4004419. DOI: 10.1186/1472-6750-14-29. View

15.

McHugh M . Interrater reliability: the kappa statistic. Biochem Med (Zagreb). 2012; 22(3):276-82. PMC: 3900052. View

16.

Anderson D, Kaye K, Classen D, Arias K, Podgorny K, Burstin H . Strategies to prevent surgical site infections in acute care hospitals. Infect Control Hosp Epidemiol. 2008; 29 Suppl 1:S51-61. DOI: 10.1086/591064. View

17.

Soroka M, Wasowicz B, Rymaszewska A . Loop-Mediated Isothermal Amplification (LAMP): The Better Sibling of PCR?. Cells. 2021; 10(8). PMC: 8393631. DOI: 10.3390/cells10081931. View

18.

Anupama K, Nayak A, Karunasagar I, Maiti B . Rapid visual detection of Vibrio parahaemolyticus in seafood samples by loop-mediated isothermal amplification with hydroxynaphthol blue dye. World J Microbiol Biotechnol. 2020; 36(5):76. DOI: 10.1007/s11274-020-02851-0. View

19.

Metwally L, Gomaa N, Hassan R . Detection of methicillin-resistant Staphylococcus aureus directly by loop-mediated isothermal amplification and direct cefoxitin disk diffusion tests. East Mediterr Health J. 2014; 20(4):273-9. View

20.

Koide Y, Maeda H, Yamabe K, Naruishi K, Yamamoto T, Kokeguchi S . Rapid detection of mecA and spa by the loop-mediated isothermal amplification (LAMP) method. Lett Appl Microbiol. 2010; 50(4):386-92. DOI: 10.1111/j.1472-765X.2010.02806.x. View