Optimization of a Quantitative PCR Methodology for Detection of Aspergillus spp. and Rhizopus arrhizus.

ABSTRACT:

Introduction

Multiplex quantitative polymerase chain reaction (qPCR) methods for the detection of Aspergillus spp. based only on SYBR Green and melting curve analysis of PCR products are difficult to develop because most targets are located within ITS regions. The aim of this study was to adapt our previously developed methodology based on a multiplex PCR assay coupled with GeneScan analysis to provide a qPCR method.

Methods

A SYBR Green-based real-time PCR assay was optimized to detect A. fumigatus, A. flavus, A. niger, A. terreus, and R. arrhizus in a multiplex assay and applied to cultured fungi and spiked plasma.

Results

Different melting temperatures allowed identification of all five pathogens and discrimination between them, even in samples with low amounts of

SUBMITTER: Mendonca A 

PROVIDER: S-EPMC9202985 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature