Direct detection of methicillin-resistant in Staphylococcus spp. in positive blood culture by isothermal recombinase polymerase amplification combined with lateral flow dipstick assay

Srisrattakarn, Arpasiri; Tippayawat, Patcharaporn; Chanawong, Aroonwadee; Tavichakorntrakool, Ratree; Daduang, Jureerut; Wonglakorn, Lumyai; Sooksongsoontorn, Pirom; Lulitanond, Aroonlug

doi:10.1007/s11274-020-02938-8

Cited by 15 publications

(29 citation statements)

References 44 publications

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“…The RPA-LF method has been adapted to detect many pathogens which requires minimal equipment and a quick turnaround time of 30 min. It has been applied for detection of pathogens in various samples, such as blood ( Srisrattakarn et al, 2020 ), plasma ( Qi et al, 2018 ), foodstuffs ( Du et al, 2018a ; Du et al, 2018b ), stool ( Crannell et al, 2014a ) and environmental samples such as plant ( Londono, Harmon & Polston, 2016 ), soil and water ( Saxena et al, 2019 ). To the best of our knowledge, this is the first report of the application of the RPA-LF method for direct detection of E .…”

Section: Discussionmentioning

confidence: 99%

“…faecium and vanA and vanB genes) were 37 °C for 20 min. Several investigations have shown that the RPA-LF can function well at temperatures ranging from 25–45 °C ( Lu et al, 2021 ; Peng et al, 2019 ) and the faint band was visible in 5 min ( Du et al, 2018a ; Peng et al, 2019 ; Srisrattakarn et al, 2020 ; Wu et al, 2020 ). Similarly, our study found that the RPA worked effectively at a wide temperature range (25–42 °C) and incubation times of 5-30 min could generate a visual signal on the test lines.…”

Section: Discussionmentioning

confidence: 99%

“…The RPA-AGE reaction was performed using TwistAmp Basic kit (TwistDX, Cambridge, UK). RPA components were mixed according to a previous study ( Srisrattakarn et al, 2020 ). The RPA reaction was incubated at 37 °C for 20 min and the reaction was stopped by heating at 65 °C for 10 min to eliminate the excess protein ( Kapoor et al, 2017 ).…”

Section: Methodsmentioning

confidence: 99%

“…Each of the 126 bacterial isolates was prepared to a suspension of 10 5 CFU/mL in normal saline solution. Spiked blood-culture samples were prepared by mixing 500 µL of the bacterial suspension with 10 mL of healthy human blood and transferred to pre-incubated aerobic culture bottles (RENDER; Zhuhai Meihua Medical Technology Limited, Zhuhai, China) to make a final inoculum of approximately 500 CFU/mL ( Srisrattakarn et al, 2020 ) and incubated overnight at 37 °C. The number of bacteria after positive signal were approximately 10 8 –10 9 CFU/mL ( Smith, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

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Rapid detection ofEnterococcusand vancomycin resistance using recombinase polymerase amplification

Panpru

Srisrattakarn

Panthasri

et al. 2021

PeerJ

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Vancomycin-resistant enterococci (VRE), especially Enterococcus faecium, have been a global concern, often causing serious healthcare-associated infections. We established a rapid approach for detecting E. faecium and vancomycin-resistance genes (vanA and vanB) in clinical samples using isothermal recombinase polymerase amplification (RPA) combined with a lateral-flow (LF) strip. Specific RPA primer sets and probes for ddl (to identify the presence of E. faecium) vanA and vanB genes were designed. The RPA reaction was performed under isothermal condition at 37 °C within 20 min and read using the LF strip within a further 5 min. A total of 141 positive blood-cultures and 136 stool/rectal swab samples were tested using RPA-LF method compared to the conventional PCR method. The RPA-LF method exhibited 100% sensitivity in both blood-culture (60 E. faecium; 35 vanA type and two vanB type) and stool/rectal-swab samples (63 E. faecium and 36 vanA type) without cross-reaction (100% specificity). The lower detection limit of the RPA-LF was approximately 10 times better than that of the conventional PCR method. The RPA-LF method is an alternative rapid method with excellent sensitivity and specificity for detecting E. faecium, vanA, and vanB, and it has the potential to be used as a point-of-care device for VRE therapy and prevention.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Rapid detection ofEnterococcusand vancomycin resistance using recombinase polymerase amplification

Panpru

Srisrattakarn

Panthasri

et al. 2021

PeerJ

Self Cite

View full text Add to dashboard Cite

show abstract

“…In Figure 11d, the parts of a LF strip are depicted. Due to their simplicity of use and low cost, these type of assays have been combined with different types of isothermal methods such as LAMP, RPA, or SEA for naked-eye detection of different microorganisms in foods, surfaces, and even clinical samples (Azinheiro et al, 2020;Du et al, 2018;Wang et al, 2020;Zasada et al, 2020), with particular interest in low-resource settings due to the need of very little instrumentation (Mei et al, 2019;Poulton & Webster, 2018;Srisrattakarn et al, 2020;van Dam et al, 2013).…”

Section: Lateral Flowmentioning

confidence: 99%

Naked‐eye detection strategies coupled with isothermal nucleic acid amplification techniques for the detection of human pathogens

Garrido‐Maestu

Prado

2022

Comp Rev Food Sci Food Safe

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Nucleic acid amplification‐based techniques have gained acceptance by the scientific, and general, community as reference methodologies for many different applications. Since the development of the gold standard of these techniques, polymerase chain reaction (PCR), back in the 1980s many improvements have been made, and alternative techniques emerged reporting improvements over PCR. Among these, isothermal amplification approaches resulted of particular interest as could overcome the need of specialized equipment to accurately control temperature changes, but it was after year 2000 that these techniques have flourished in a huge number of novel alternatives with many different degrees of complexities and requirements. An added value is their possibility to be combined with many different naked‐eye detection strategies, simplifying the resources needed, allowing to reduce cost, and serving as the basis for novel developments of lab‐on‐chip systems, and miniaturized devices, for point‐of‐care testing. In this review, we will go over different types of naked‐eye detection strategies, combined with isothermal amplification. This will provide the readers up‐to‐date information for them to select the most appropriate strategies depending on the particular needs and resources for their experimental setup.

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Development of Two Recombinase Polymerase Amplification EXO (RPA-EXO) and Lateral Flow Dipstick (RPA-LFD) Techniques for the Rapid Visual Detection of Aeromonas salmonicida

Zhou¹,

Zheng²,

Yang³

et al. 2022

Mar Biotechnol

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Direct detection of methicillin-resistant in Staphylococcus spp. in positive blood culture by isothermal recombinase polymerase amplification combined with lateral flow dipstick assay

Cited by 15 publications

References 44 publications

Rapid detection ofEnterococcusand vancomycin resistance using recombinase polymerase amplification

Rapid detection ofEnterococcusand vancomycin resistance using recombinase polymerase amplification

Naked‐eye detection strategies coupled with isothermal nucleic acid amplification techniques for the detection of human pathogens

Development of Two Recombinase Polymerase Amplification EXO (RPA-EXO) and Lateral Flow Dipstick (RPA-LFD) Techniques for the Rapid Visual Detection of Aeromonas salmonicida

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