Feasibility of Loop-Mediated Isothermal Amplification for Rapid Detection of Methicillin-Susceptible and Methicillin-Resistant Staphylococcus aureus in Tissue Samples

Sang Gyun Kim; Gi Won Choi; Won Seok Choi; Chae Seung Lim; Woong Sik Jang; Ji Hoon Bae

doi:10.4055/cios21277

Feasibility of Loop-Mediated Isothermal Amplification for Rapid Detection of Methicillin-Susceptible and Methicillin-Resistant Staphylococcus aureus in Tissue Samples

Sang Gyun Kim, Gi Won Choi, Won Seok Choi, Chae Seung Lim, Woong Sik Jang, Ji Hoon Bae

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

Abstract

Background: To date, few studies have investigated the feasibility of the loop-mediated isothermal amplification (LAMP) assay for identifying pathogens in tissue samples. This study aimed to investigate the feasibility of LAMP for the rapid detection of methicillin-susceptible or methicillin-resistant Staphylococcus aureus (MSSA or MRSA) in tissue samples, using a bead-beating DNA extraction method. Methods: Twenty tissue samples infected with either MSSA (n = 10) or MRSA (n = 10) were obtained from patients who underwent orthopedic surgery for suspected musculoskeletal infection between December 2019 and September 2020. DNA was extract-ed from the infected tissue samples using the bead-beating method. A multiplex LAMP assay was conducted to identify MSSA and MRSA infections. To recognize the Staphylococcus genus, S. aureus, and methicillin resistance, 3 sets of 6 primers for the 16S ribosomal ribonucleic acid (rRNA) and the femA and mecA genes were used, respectively. The limit of detection and sensitivity (de-tection rate) of the LAMP assay for diagnosing MSSA and MRSA infection were analyzed. Results: The LAMP result was positive for samples containing 10³ colony-forming unit (CFU)/mL for 16S rRNA, 10⁴ CFU/mL for femA, and 10⁵ CFU/mL for mecA. The limits of detection for 16S rRNA and femA were not different between MSSA and MRSA. For the 10 MSSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and femA and a 100% negative reaction for mecA. For the 10 MRSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and mecA but only 90% positive reactions for femA. The sensitivity (detection rate) of the LAMP assay for identifying MSSA and MRSA in infected tissue samples was 100% and 90%, respectively. Conclusions: The results of this study suggest that the LAMP assay performed with tissue DNA samples can be a useful diagnostic method for the rapid detection of musculoskeletal infections caused by MSSA and MRSA.

Original language	English
Pages (from-to)	466-473
Number of pages	8
Journal	CiOS Clinics in Orthopedic Surgery
Volume	14
Issue number	3
DOIs	https://doi.org/10.4055/cios21277
Publication status	Published - 2022 Sept 1
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2022 by The Korean Orthopaedic Association.

Keywords

Loop-mediated isothermal amplification
Methicillin-resistant Staphylococcus aureus
Methicillin-susceptible Staphylococcus aureus

ASJC Scopus subject areas

Surgery
Orthopedics and Sports Medicine

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10.4055/cios21277

Cite this

@article{b2592a2a4c274501834279fa5f80efe2,

title = "Feasibility of Loop-Mediated Isothermal Amplification for Rapid Detection of Methicillin-Susceptible and Methicillin-Resistant Staphylococcus aureus in Tissue Samples",

abstract = "Background: To date, few studies have investigated the feasibility of the loop-mediated isothermal amplification (LAMP) assay for identifying pathogens in tissue samples. This study aimed to investigate the feasibility of LAMP for the rapid detection of methicillin-susceptible or methicillin-resistant Staphylococcus aureus (MSSA or MRSA) in tissue samples, using a bead-beating DNA extraction method. Methods: Twenty tissue samples infected with either MSSA (n = 10) or MRSA (n = 10) were obtained from patients who underwent orthopedic surgery for suspected musculoskeletal infection between December 2019 and September 2020. DNA was extract-ed from the infected tissue samples using the bead-beating method. A multiplex LAMP assay was conducted to identify MSSA and MRSA infections. To recognize the Staphylococcus genus, S. aureus, and methicillin resistance, 3 sets of 6 primers for the 16S ribosomal ribonucleic acid (rRNA) and the femA and mecA genes were used, respectively. The limit of detection and sensitivity (de-tection rate) of the LAMP assay for diagnosing MSSA and MRSA infection were analyzed. Results: The LAMP result was positive for samples containing 103 colony-forming unit (CFU)/mL for 16S rRNA, 104 CFU/mL for femA, and 105 CFU/mL for mecA. The limits of detection for 16S rRNA and femA were not different between MSSA and MRSA. For the 10 MSSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and femA and a 100% negative reaction for mecA. For the 10 MRSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and mecA but only 90% positive reactions for femA. The sensitivity (detection rate) of the LAMP assay for identifying MSSA and MRSA in infected tissue samples was 100% and 90%, respectively. Conclusions: The results of this study suggest that the LAMP assay performed with tissue DNA samples can be a useful diagnostic method for the rapid detection of musculoskeletal infections caused by MSSA and MRSA.",

keywords = "Loop-mediated isothermal amplification, Methicillin-resistant Staphylococcus aureus, Methicillin-susceptible Staphylococcus aureus",

author = "Kim, {Sang Gyun} and Choi, {Gi Won} and Choi, {Won Seok} and Lim, {Chae Seung} and Jang, {Woong Sik} and Bae, {Ji Hoon}",

note = "Publisher Copyright: {\textcopyright} 2022 by The Korean Orthopaedic Association.",

year = "2022",

month = sep,

day = "1",

doi = "10.4055/cios21277",

language = "English",

volume = "14",

pages = "466--473",

journal = "CiOS Clinics in Orthopedic Surgery",

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T1 - Feasibility of Loop-Mediated Isothermal Amplification for Rapid Detection of Methicillin-Susceptible and Methicillin-Resistant Staphylococcus aureus in Tissue Samples

AU - Kim, Sang Gyun

AU - Choi, Gi Won

AU - Choi, Won Seok

AU - Lim, Chae Seung

AU - Jang, Woong Sik

AU - Bae, Ji Hoon

PY - 2022/9/1

Y1 - 2022/9/1

N2 - Background: To date, few studies have investigated the feasibility of the loop-mediated isothermal amplification (LAMP) assay for identifying pathogens in tissue samples. This study aimed to investigate the feasibility of LAMP for the rapid detection of methicillin-susceptible or methicillin-resistant Staphylococcus aureus (MSSA or MRSA) in tissue samples, using a bead-beating DNA extraction method. Methods: Twenty tissue samples infected with either MSSA (n = 10) or MRSA (n = 10) were obtained from patients who underwent orthopedic surgery for suspected musculoskeletal infection between December 2019 and September 2020. DNA was extract-ed from the infected tissue samples using the bead-beating method. A multiplex LAMP assay was conducted to identify MSSA and MRSA infections. To recognize the Staphylococcus genus, S. aureus, and methicillin resistance, 3 sets of 6 primers for the 16S ribosomal ribonucleic acid (rRNA) and the femA and mecA genes were used, respectively. The limit of detection and sensitivity (de-tection rate) of the LAMP assay for diagnosing MSSA and MRSA infection were analyzed. Results: The LAMP result was positive for samples containing 103 colony-forming unit (CFU)/mL for 16S rRNA, 104 CFU/mL for femA, and 105 CFU/mL for mecA. The limits of detection for 16S rRNA and femA were not different between MSSA and MRSA. For the 10 MSSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and femA and a 100% negative reaction for mecA. For the 10 MRSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and mecA but only 90% positive reactions for femA. The sensitivity (detection rate) of the LAMP assay for identifying MSSA and MRSA in infected tissue samples was 100% and 90%, respectively. Conclusions: The results of this study suggest that the LAMP assay performed with tissue DNA samples can be a useful diagnostic method for the rapid detection of musculoskeletal infections caused by MSSA and MRSA.

AB - Background: To date, few studies have investigated the feasibility of the loop-mediated isothermal amplification (LAMP) assay for identifying pathogens in tissue samples. This study aimed to investigate the feasibility of LAMP for the rapid detection of methicillin-susceptible or methicillin-resistant Staphylococcus aureus (MSSA or MRSA) in tissue samples, using a bead-beating DNA extraction method. Methods: Twenty tissue samples infected with either MSSA (n = 10) or MRSA (n = 10) were obtained from patients who underwent orthopedic surgery for suspected musculoskeletal infection between December 2019 and September 2020. DNA was extract-ed from the infected tissue samples using the bead-beating method. A multiplex LAMP assay was conducted to identify MSSA and MRSA infections. To recognize the Staphylococcus genus, S. aureus, and methicillin resistance, 3 sets of 6 primers for the 16S ribosomal ribonucleic acid (rRNA) and the femA and mecA genes were used, respectively. The limit of detection and sensitivity (de-tection rate) of the LAMP assay for diagnosing MSSA and MRSA infection were analyzed. Results: The LAMP result was positive for samples containing 103 colony-forming unit (CFU)/mL for 16S rRNA, 104 CFU/mL for femA, and 105 CFU/mL for mecA. The limits of detection for 16S rRNA and femA were not different between MSSA and MRSA. For the 10 MSSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and femA and a 100% negative reaction for mecA. For the 10 MRSA-positive samples, the LAMP assay showed 100% positive reactions for 16S rRNA and mecA but only 90% positive reactions for femA. The sensitivity (detection rate) of the LAMP assay for identifying MSSA and MRSA in infected tissue samples was 100% and 90%, respectively. Conclusions: The results of this study suggest that the LAMP assay performed with tissue DNA samples can be a useful diagnostic method for the rapid detection of musculoskeletal infections caused by MSSA and MRSA.

KW - Loop-mediated isothermal amplification

KW - Methicillin-resistant Staphylococcus aureus

KW - Methicillin-susceptible Staphylococcus aureus

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JO - CiOS Clinics in Orthopedic Surgery

JF - CiOS Clinics in Orthopedic Surgery

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ER -

Feasibility of Loop-Mediated Isothermal Amplification for Rapid Detection of Methicillin-Susceptible and Methicillin-Resistant Staphylococcus aureus in Tissue Samples

Abstract

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Keywords

ASJC Scopus subject areas

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