Optimizing bacterial DNA extraction in urine.

08:00 EDT 24th September 2019 | BioPortfolio

Summary of "Optimizing bacterial DNA extraction in urine."

Urine is an acceptable, non-invasive sample for investigating the human urogenital microbiota and for the diagnosis of sexually transmitted infections. However, low quantities of bacterial DNA and PCR inhibitors in urine may prevent efficient PCR amplification for molecular detection of bacteria. Furthermore, cold temperatures used to preserve DNA and bacteria in urine can promote precipitation of crystals that interfere with DNA extraction. Saline, Dulbecco's Phosphate Buffered Saline, or Tris-EDTA buffer were added to urine from adult men to determine if crystal precipitation could be reversed without heating samples beyond ambient temperature. Total bacterial DNA concentrations and PCR inhibition were measured using quantitative PCR assays to compare DNA yields with and without buffer addition. Dissolution of crystals with Tris-EDTA prior to urine centrifugation was most effective in increasing bacterial DNA recovery and reducing PCR inhibition. DNA recovery using Tris-EDTA was further tested by spiking urine with DNA from bacterial isolates and median concentrations of Lactobacillus jensenii and Escherichia coli 16S rRNA gene copies were found to be higher in urine processed with Tris-EDTA. Maximizing bacterial DNA yield from urine may facilitate more accurate assessment of bacterial populations and increase detection of specific bacteria in the genital tract.


Journal Details

This article was published in the following journal.

Name: PloS one
ISSN: 1932-6203
Pages: e0222962


DeepDyve research library

PubMed Articles [7028 Associated PubMed Articles listed on BioPortfolio]

Developed nano carbon-based coating for simultaneous extraction of potent central nervous system stimulants from urine media by stir bar sorptive extraction method coupled to high performance liquid chromatography.

A nano graphene oxide sol-gel composite (NGO/sol-gel) applied as a coating of a capillary glass tube stir bar to develop a novel stir bar sorptive extraction (SBSE) method for simultaneous extraction ...

Optimization of dispersive liquid-liquid microextraction procedure for detecting chlorpyrifos in human urine samples.

Selecting an effective sample preparation method to measure target pesticides in biological matrices is a serious challenge for researchers. This study aimed to optimize the dispersive liquid-liquid m...

A pharmacokinetics study of orally administered higenamine in rats using LC-MS/MS for doping control analysis.

According to WADA guidelines, the presence of Higenamine (HG) in urine should not be ≥ 10 ng/mL. HG is widely found in materials used in Chinese herbal medicines as well as food and additives. This ...

Analytical Comparison of Methods for Extraction of Short Cell-Free DNA from Urine.

Urine cell-free DNA (cfDNA) is a valuable noninvasive biomarker for detecting cancer mutation, diagnosing infectious disease (eg, tuberculosis), monitoring organ transplantation, and prenatal screenin...

Selective extraction and detection of β-agonists in swine urine for monitoring illegal use in livestock breeding.

The illegal use of β-agonists often endangers animal-derived food safety. In this study, a selective detection method for β-agonists in swine urine was established via the combination of polymeric i...

Clinical Trials [3921 Associated Clinical Trials listed on BioPortfolio]

Changes in Resistome After Dental Extraction and Amoxicillin.

The aim of this study is to evaluate the nature of the change in oral and gut bacterial resistance profiles after antibiotic therapy for a surgical procedure in two groups. The interventio...

The Effects of Cranberry Juice on Bacterial Adhesion

The study is based on the hypothesis that consumption of cranberry juice products results in the production of metabolites in the urine that have anti-adhesive properties on bacteria. A cr...

Evaluation of Urine Samples Obtained by Bladder Stimulation for the Diagnosis of Urinary Tract Infection in Infants

Urinary tract infection (UTI) is the most common serious bacterial infection among infants. Suprapubic aspiration and bladder catheterization are considered as the gold standard by the Ame...

Evaluation of Post-operative Discomforts in Children After Extraction of Temporary Teeth

This prospective observational study aim to evaluate discomforts after extraction of temporary tooth under local anesthesia. The first objective is to describe the prevalence of the post-e...

3D Radiographic Evaluation of a Novel Implant Site Development Technique

The present retrospective study aims at assessing the clinical effects of a novel implant site development (Extraction site development - XSD) technique after tooth extraction compared wit...

Medical and Biotech [MESH] Definitions

Use of various chemical separation and extraction methods, such as SOLID PHASE EXTRACTION; CHROMATOGRAPHY; and SUPERCRITICAL FLUID EXTRACTION; to prepare samples for analytical measurement of components.

The selective extraction of deciduous teeth during the stage of mixed dentition in accordance with the shedding and eruption of the teeth. It is done over an extended period to allow autonomous adjustment to relieve crowding of the dental arches during the eruption of the lateral incisors, canines, and premolars, eventually involving the extraction of the first premolar teeth. (Dorland, 28th ed)

Extraction of the fetus by means of obstetrical instruments.

An extraction method that separates analytes using a solid phase and a liquid phase. It is used for preparative sample cleanup before analysis by CHROMATOGRAPHY and other analytical methods.

A condition sometimes occurring after tooth extraction, particularly after traumatic extraction, resulting in a dry appearance of the exposed bone in the socket, due to disintegration or loss of the blood clot. It is basically a focal osteomyelitis without suppuration and is accompanied by severe pain (alveolalgia) and foul odor. (Dorland, 28th ed)

Quick Search

DeepDyve research library

Relevant Topics

DNA Extraction
DNA is extracted from human cells for a variety of reasons. With a pure sample of DNA you can test a newborn for a genetic disease, analyze forensic evidence, or study a gene involved in cancer. DNA isolation is a process of purification of DNA from...

Benign Prostatic Hyperplasia (BPH) Erectile Dysfunction Urology Urology is the branch of medicine concerned with the urinary tract and diseases that affect it. Examples include urethritis, urethrostenosis and incontinence. Urology is a su...

Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...

Searches Linking to this Article