Multiplex Digital PCR-Based Development and discussion of the Detection of Genetic Association Between Staphylococcus aureus and mecA

Abstract

Ciming Wu,1,* Ming Ling,2,* Qiong Chen,3 Hui Chai,1 Huan Chen1,4 1Zhejiang Chinese Medical University, College of Life Science, Hangzhou, Zhejiang, 310012, People’s Republic of China; 2Jinhua Institute for Food and Drug Control, Jinhua, Zhejiang, People’s Republic of China; 3Hangzhou First People’s Hospital, Hangzhou, Zhejiang, People’s Republic of China; 4Hangzhou Digital-Micro Biotech Co, Ltd, Hangzhou, Zhejiang, People’s Republic of China*These authors contributed equally to this workCorrespondence: Huan Chen; Hui Chai, Email [email protected]; [email protected]: Methicillin-resistant Staphylococcus aureus (MRSA) is a predominant nosocomial infection-causing bacteria. The aim of this study was to develop a novel single-bacteria multiplex digital PCR assays (SMD-PCR), which is capable of simultaneously detecting and discriminating Methicillin-sensitive Staphylococcus aureus (MSSA) and MRSA. This protocol employed TaqMan probes to detect SAOUHSC_00106 and mecA genes, with the latter being linked to methicillin resistance. A total of 72 samples from various specimen types were evaluated. The accuracy rates for the sputum samples, pus samples, swab samples, ear secretion samples, and catheter samples were 94.44%, 100%, 92%, 100%, and 100%, respectively. Our results showed that the clinical practicability of SMD-PCR has applicability to the rapid detection of MRSA without DNA extraction or bacterial culture, and can be utilized for the rapid detection of Staphylococcus aureus and the timely identification of MRSA in clinical samples, thereby providing an advanced platform for the rapid diagnosis of clinical MRSA infection.Keywords: MSSA, MRSA, droplet digital PCR, SMD-PCR

Keywords

• mssa
• mrsa
• droplet digital pcr
• smd-pcr