A novel approach to eliminate detection of contaminating Staphylococcal species introduced during clinical testing

Ao, Wanyuan; Clifford, Adrianne; Corpuz, Maylene; Jenison, Robert

doi:10.1371/journal.pone.0171915

Cited by 3 publications

(2 citation statements)

References 24 publications

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“…Elimination of unintended contamination during sample processing is of utmost importance in clinical diagnosis. Addition of a non-target specific sequence at a large copy number or contaminating the sample with a defined amount of bacteria, the amplification of the contaminant species can be completely suppressed because of the limited amount of HDA primers [71]. It is important that the nontarget sequences/organisms have some level of homology with the target to consume the primers.…”

Section: Applicationsmentioning

confidence: 99%

Helicase-dependent isothermal amplification: a novel tool in the development of molecular-based analytical systems for rapid pathogen detection

Barreda-García

Miranda‐Castro

de‐los‐Santos‐Álvarez

et al. 2017

Anal Bioanal Chem

111

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Highly sensitive testing of nucleic acids is essential to improve the detection of pathogens, which pose a major threat for public health worldwide. Currently available molecular assays, mainly based on PCR, have a limited utility in point-of-need control or resource-limited settings. Consequently, there is a strong interest in developing cost-effective, robust, and portable platforms for early detection of these harmful microorganisms. Since its description in 2004, isothermal helicase-dependent amplification (HDA) has been successfully applied in the development of novel molecular-based technologies for rapid, sensitive, and selective detection of viruses and bacteria. In this review, we highlight relevant analytical systems using this simple nucleic acid amplification methodology that takes place at a constant temperature and that is readily compatible with microfluidic technologies. Different strategies for monitoring HDA amplification products are described. In addition, we present technological advances for integrating sample preparation, HDA amplification, and detection. Future perspectives and challenges toward point-of-need use not only for clinical diagnosis but also in food safety testing and environmental monitoring are also discussed. Graphical Abstract Expanding the analytical toolbox for the detection of DNA sequences specific of pathogens with isothermal helicase dependent amplification (HDA).

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Section: Applicationsmentioning

confidence: 99%

Helicase-dependent isothermal amplification: a novel tool in the development of molecular-based analytical systems for rapid pathogen detection

Barreda-García

Miranda‐Castro

de‐los‐Santos‐Álvarez

et al. 2017

Anal Bioanal Chem

111

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“…114 Careful screening of vendors and using methods that remove or suppress contaminations are often required to achieve robust PCR performance, especially for detecting low-abundance targets. 115 Finally, PCR assays are typically designed to detect a limited set of preidentified genes, which are not able to cover rapid and complex evolving mechanisms associated with infectious bacteria. In particular, it remains a challenge for conventional PCR to detect many of the ESBLs and CREs that are highly variable and often differ from each other by single-nucleotide polymorphisms (SNPs).…”

Section: Emerging Microtechnologies and Automated Systems For Moleculmentioning

confidence: 99%

Emerging Microtechnologies and Automated Systems for Rapid Bacterial Identification and Antibiotic Susceptibility Testing

Xing²,

Zhao

2017

SLAS Technology

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Rapid bacterial identification (ID) and antibiotic susceptibility testing (AST) are in great demand due to the rise of drug-resistant bacteria. Conventional culture-based AST methods suffer from a long turnaround time. By necessity, physicians often have to treat patients empirically with antibiotics, which has led to an inappropriate use of antibiotics, an elevated mortality rate and healthcare costs, and antibiotic resistance. Recent advances in miniaturization and automation provide promising solutions for rapid bacterial ID/AST profiling, which will potentially make a significant impact in the clinical management of infectious diseases and antibiotic stewardship in the coming years. In this review, we summarize and analyze representative emerging micro- and nanotechnologies, as well as automated systems for bacterial ID/AST, including both phenotypic (e.g., microfluidic-based bacterial culture, and digital imaging of single cells) and molecular (e.g., multiplex PCR, hybridization probes, nanoparticles, synthetic biology tools, mass spectrometry, and sequencing technologies) methods. We also discuss representative point-of-care (POC) systems that integrate sample processing, fluid handling, and detection for rapid bacterial ID/AST. Finally, we highlight major remaining challenges and discuss potential future endeavors toward improving clinical outcomes with rapid bacterial ID/AST technologies.

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Antibiotic susceptibility and phenotypic profile of Staphylococcus species isolated from different clinical samples from health facilities: A cross-sectional study

Anwar,

Saadalla,

Muhammad Amin

et al. 2024

SAGE Open Medicine

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Background: Staphylococcus species are widely distributed in nature and found in various human body sites. Objectives: To determine the antibiotic susceptibility pattern of Staphylococcus species isolated from different clinical samples. Methods: This cross-sectional study was conducted on 400 clinical specimens from conveniently sampled patients seeking healthcare at two health facilities in sulaimani / Iraq. Bacterial isolation and identification were done using conventional techniques, after which the antibiotic susceptibility profile of Staphylococcus species commonly prescribed antibiotics used in treating infections at the facilities was done using the disc diffusion method. Finally, MecA, methicillin-resistant Staphylococcus aureus and macrolides-lincosamide and streptogramin genes with mupirocin-resistant, beta-lactamase and vancomycin-resistance phenotypes were identified. Results: Staphylococcus aureus was the prevalent isolated species ( n = 197, 49.3%), followed by Staphylococcus hemolyticus ( n = 115, 28.8%), Staphylococcus epidermidis ( n = 49, 12.3%), Staphylococcus hominis ( n = 9.0, 2.3%), Staphylococcus sciuri ( n = 8.0, 2.0%) and Staphylococcus lentus ( n = 4.0, 1.0%). All isolated species resisted Penicillin G, Ampicillin, Cefotaxime and Cefoxitin. Most of the isolates, 89.5% ( n = 358) had the beta-lactamase phenotype, 18.0% ( n = 72) had the MecA gene, 2.8% ( n = 11) the Mupirocin-resistant phenotype, and 2.0% ( n = 8.0) the vancomycin-resistance phenotype. Additionally, 12 isolates had both methicillin-resistant Staphylococcus aureus (66.7%) and macrolides-lincosamide and streptogramin (65.2%) genes. The majority of the patients, 43% ( n = 172) were >50 years old and 52.25% ( n = 209) males. Also, most samples were from patients with urinary tract infection ( n = 73), wound ( n = 71), blood ( n = 35), sputum ( n = 29), pus ( n = 28), seminal fluid ( n = 27), cerebrospinal fluid ( n = 1.0) and stool ( n = 1.0). Most isolates that had the MSLb gene were highly significantly resistant to both Clindamycin (94.6%) and Erythromycin (84.7%) ( p < 0.001). Conclusions: Staphylococcus aureus was the predominant Staphylococcus species isolated from the clinical samples, most of which were resistant to most commonly prescribed antibiotics and had developed resistant genes and phenotypes.

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A novel approach to eliminate detection of contaminating Staphylococcal species introduced during clinical testing

Cited by 3 publications

References 24 publications

Helicase-dependent isothermal amplification: a novel tool in the development of molecular-based analytical systems for rapid pathogen detection

Helicase-dependent isothermal amplification: a novel tool in the development of molecular-based analytical systems for rapid pathogen detection

Emerging Microtechnologies and Automated Systems for Rapid Bacterial Identification and Antibiotic Susceptibility Testing

Antibiotic susceptibility and phenotypic profile of Staphylococcus species isolated from different clinical samples from health facilities: A cross-sectional study

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