Modified PCR protocol to increase sensitivity for determination of bacterial community composition.

ABSTRACT:

Background

The objective of this project was to increase the sensitivity of sequence-based bacterial community determination without impacting community composition or interfering with cluster formation during sequencing. Two PCR protocols (standard and modified) were examined in airway samples where we observed a large range in bacterial load (3.1-6.2 log₁₀ 16S rRNA gene copies/reaction). Tracheal aspirate (TA) samples (n = 99) were collected from sixteen children requiring mechanical ventilation at a single center. DNA was extracted, and total bacterial load (TBL) was assessed using qPCR. Amplification of 16S rRNA was attempted with both protocols in all samples.

Results

PCR product was observed using both protocols in 52 samples and in 24 additional samples onl

SUBMITTER: Williamson KM 

PROVIDER: S-EPMC8045227 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature