Molecular methods for biofilms

Ferrera, Isabel; Balagué, Vanessa; Voolstra, Christian R.; Aranda, Manuel; Bayer, Till; Abed, Raeid M. M.; Dobretsov, Sergey; Owens, Sarah M.; Wilkening, Jared; Fessler, Jennifer L.; Gilbert, Jack A.

doi:10.1002/9781118336144.ch4

Cited by 1 publication

(1 citation statement)

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“…The development of molecular approaches that are able to identify many species in parallel by examining the genetic content or microbiome provides a more comprehensive view of microbial communities (11). Several indirect methods, including fingerprinting [denaturing gradient gel electrophoresis (DGGE), terminal restriction fragment length polymorphisms (T-RFLP), temperature-time gradient gel electrophoresis (TTGE)], hybridization (clonal hybridization, microarray), and PCR Multiple touch has been used to investigate the microbiota in clinical samples (12). The aim of this study was to develop a multiplex PCR method to detect H. influenzae, S. pneumoniae and S. pyogenes in clinical sputum samples.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Genetically Detection of Streptococcus pyogenes, Streptococcus pneumoniae and Haemophilus influenzae in Patients with Treatment-Resistant Respiratory Infection

Mohammadi Ebli,

Heshmatipour,

Daneshjou

et al. 2023

Iran J Pathol

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Background & Objective: Streptococcus pneumoniae , Haemophilus influenzae and Streptococcus pyogenes are among the most important causes of infection in human. Inventing rapid methods to identify these species can help in providing appropriate and effective treatment options. Therefore, the current study aimed to develop a multiplex touch-down PCR method to identify rapidly the aforementioned species patients' sputum samples, simultaneously. Methods: A total of 50 sputum samples of patients with respiratory infections resistant to treatment were collected. After DNA extraction and primer design, the complete capsule (CAP) region II, capsular polysaccharide biosynthesis (cpsA) and the structural regulator of transcription (spy) genes were amplified for detecting H. influenzae , S. pneumoniae and S. pyogenes by multiplex touch-down PCR. Results: Among 50 samples prepared from patients with different diseases, 27 samples were positive for amplified genes. The frequency of presence of pathogens in the collected samples included 14% H. influenzae , 20% S. pneumoniae and 20% S. pyogenes . Also, in some patients, the simultaneous presence of two or three pathogens were observed. Conclusion: In general, it can be concluded that the PCR touchdown method developed in the present study is an effective and fast method for the simultaneous identification of H. influenzae , S. pneumoniae and S. pyogenes pathogens in clinical samples of patients.

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Section: Introductionmentioning

confidence: 99%