Infection and Drug Resistance (Aug 2023)
A PCR-Reverse Dot Blot Hybridization Based Microfluidics Detection System for the Rapid Identification of 13 Fungal Pathogens Directly After Blood Cultures Over a Period of Time
Abstract
Chunhui Lin,1,2,* Hao Tang,3,* Xinyi Hu,1,2 Ge Li,1,2 Tong Jiang,1,2 Wensu Yang,1,2 Zhaoxin Xia,1,2 Yi Zhu,1,2 Huaming Xu,4 Jing Zhou,1,2 Jilu Shen1,2 1Clinical Laboratory, the First Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 2Clinical Laboratory, Anhui Public Health Clinical Center Hefei, Hefei, People’s Republic of China; 3Clinical Laboratory, the Second Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 4Clinical Laboratory, the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jilu Shen, Tel +86 151 5515 2963, Email [email protected]: It is time-consuming to identify fungal pathogens from positive blood cultures using the standard culture-based method. And delayed diagnosis of bloodstream infection leads to significantly increased mortality.Methods: We developed a PCR-reverse dot blot hybridization combined with microfluidic chip techniques to rapidly identify 13 fungal pathogens within 3– 4 h using the sample of blood cultured over a period of time.Results: We performed clinical validation using 43 blood culture-positive samples with a sensitivity of 96.7%, a specificity of 100%, and a concordance rate of 97.7%. Samples with different culture durations were evaluated using our approach, showing a detection rate of 85.2% at 16 h and 96.3% at 24 h; the platform could reach a detection limit of 103cfu/mL for the Candida spp. and 103 copies/mL for Aspergillus spp.Discussion: The detection rate of the platform is much higher than the positive rates of concurrent blood cultures. This method bears substantial clinical application potential as it incorporates the microfluidic platform with low reagent consumption, automation, and low cost (about 10 dollars).Keywords: fungemia, bloodstream infection, microfluidics, Candida auris, PCR-reverse dot blot hybridization