What is broad-range 16S rDNA PCR?

نویسندگان

  • Amani Patel
  • Kathryn A Harris
  • Felicity Fitzgerald
چکیده

Patel A, et al. Arch Dis Child Educ Pract Ed 2017;0:1–4. doi:10.1136/archdischild-2016-312049 INTRODUCTION PCRs have revolutionised the detection of bacteria in clinical samples since their widespread introduction in the 1990s. Quantitative PCR (qPCR), also known as specific PCR, involves the targeting of particular bacterial species. The technique uses specific primers (short strands of nucleic acid needed to initiate DNA replication) and fluorescent probes to allow real-time quantification of target bacterial DNA during amplification. The qPCR assay is a mainstay of microbiological diagnostics within the National Health Service (NHS). At our hospital approximately 200 qPCRs are performed per week for the investigation of bacterial infections. Although qPCR is by far the most frequently used molecular technique in bacterial diagnostics, in certain scenarios a broad-range (non-specific) 16S rDNA (ribosomal DNA) PCR is increasingly being used. Broad-range 16S rDNA PCR is also more commonly used in research settings, originally for use in detecting and identifying unusual bacterial species but now more widely used in the rapidly expanding field of microbiome research. This technique provides the initial step in the process of analysing complex microbial communities in human, zoological and even geological settings. In the future, analysis of individualised microbial communities using broad-range 16S rDNA PCR may be a key component of personalised medicine.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Empyema: the use of broad range 16S rDNA PCR for pathogen detection.

BACKGROUND An increase in the incidence of thoracic empyema in children has been reported. The causative pathogen is often unknown as pleural fluid is frequently sterile at the time of culture. The role of unusual organisms is unclear. AIMS (1) To compare the detection of organisms in pleural fluid from children with empyema using a molecular technique (16S rDNA polymerase chain reaction (PCR...

متن کامل

Development and evaluation of a novel fast broad-range 16S ribosomal DNA PCR and sequencing assay for diagnosis of bacterial infective endocarditis: multi-year experience in a large Canadian healthcare zone and a literature review

BACKGROUND The study aimed to explore the sensitivity and specificity of a novel fast 16S rDNA PCR and sequencing assay for the improved diagnosis of infective endocarditis (IE) in patients with suspected native or prosthetic heart valve (HV) infection over a multi-year period at our cardiovascular center. METHODS Sixty-eight patients were prospectively enrolled who underwent HV replacement f...

متن کامل

Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction.

OBJECTIVE To evaluate broad-range 16S ribosomal DNA (rDNA) polymerase chain reaction (PCR) as a rapid screening tool to detect bacterial contamination of stem-cell products. METHODS We performed the evaluation using whole blood spiked with serially diluted bacterial-type strains. Detection sensitivity was defined as the bacterial concentration for which all replicates were positive at each co...

متن کامل

Short communication: Evidence that microbial translocation occurs in HIV-infected children in the United Kingdom.

Microbial translocation (MT) from the gut is implicated in driving immune activation, increasing morbidity and mortality in HIV. We used bacterial 16S rDNA PCR, Sanger sequencing, and high-throughput sequencing to identify microbial DNA in the bloodstream of HIV-infected children in London, United Kingdom. Blood samples were collected from sequential children attending the HIV clinic at Great O...

متن کامل

Development of broad-range 16S rDNA PCR for use in the routine diagnostic clinical microbiology service.

The aim of the present study was to develop a broad-range PCR based on bacterial 16S rDNA for use in the routine diagnostic clinical microbiology service. The optimization and validation of the assay for use on clinical specimens from normally sterile sites is described, and preliminary results are reported on the use of the assay in the clinical diagnosis of bacterial infection in 382 paediatr...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Archives of disease in childhood. Education and practice edition

دوره 102 5  شماره 

صفحات  -

تاریخ انتشار 2017