Comparative evaluation of proteinase K and dithiothreitol as pretreatments for extracting nucleic acids from respiratory samples for multiplex PCR.

ABSTRACT:

Background

Lower respiratory tract infections are frequently caused by bacteria, and the rapid identification of pathogens is crucial for guiding treatment. Multiplex PCR(M-PCR) can detect multiple pathogens simultaneously, but mucus and other cells in lower respiratory tract samples may interfere with nucleic acid detection. In this study, we compared the effectiveness of two pretreatment methods-proteinase K(PK) and dithiothreitol(DTT)-in detecting multiple pathogens using M-PCR in bronchoalveolar lavage fluid(BALF) and sputum samples.

Methods

A total of 30 BALF samples and 20 sputum samples were collected. These samples were pretreated with PK and DTT, respectively. Bacterial structural changes and background material were examined using Gram staining. Nucleic acid purity

SUBMITTER: Chang Y 

PROVIDER: S-EPMC12016214 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature