Evaluation of Droplet Digital PCR Assay for the Diagnosis of Candidemia in Blood Samples

Abstract

Numerous studies have shown that droplet digital PCR (ddPCR) is a promising tool for the diagnosis of pathogens, especially in samples with low concentrations of pathogenic DNA. An early diagnosis of candidemia is critical for the effective treatment of patients. In this study, we evaluated the sensitivity and specificity of ddPCR assay for Candida DNA detection both in vitro by mixing fungal cells with human blood and in vivo by analyzing blood samples from infected mice and patients with suspected candidemia. The results showed that ddPCR assay could detect a minimum of 4.5 DNA copies per reaction in blood samples. ddPCR showed higher sensitivity and specificity for Candida DNA detection than traditional culture and quantitative PCR (qPCR) methods and also exhibited significantly better positive and negative predictive values than the culture and qPCR methods that were commonly used in clinical practice. Hence, our study demonstrates that ddPCR assay is a promising method for the timely diagnosis of candidemia and could be useful for monitoring the treatment of candidemia.

Performance of Multiplex PCR and β-1,3-D-Glucan Testing for the Diagnosis of Candidemia

“…However, new techniques and noteworthy developments have been recently reported. The use of digital droplet PCR (ddPCR) promises higher sensitivities and specificities [44]. The T2Candida assay, a combination of nucleic acid amplification and magnetic resonance readout, allows skipping nucleic acid extraction, thereby reducing turnaround time.…”

Biological Activity and Sterilization Mechanism of Marine Fungi‐derived Aromatic Butenolide Asperbutenolide A Against Staphylococcus aureus

“…And the stander stains of candida.albicans and candida.tropical were from our lab. The strains of C. albicans and C. tropical were inoculated on Sabouraud‐glucose‐agar (SDA) plates at 30 °C as our previously protocol [23] …”

“…The strains of C. albicans and C. tropical were inoculated on Sabouraudglucose-agar (SDA) plates at 30 °C as our previously protocol. [23]…”

Advances in droplet digital polymerase chain reaction on microfluidic chips

“…135 For bacterial detection in human samples, ddPCR can be used directly for rapid taxonomic identification and antibiotic susceptibility analysis of bacteria in human whole blood samples, especially in blood samples with low bacterial DNA content, eliminating the need for processes such as bacterial culture and pre-treatment, greatly reducing experimental time and playing an important role in early diagnosis and treatment monitoring against bacterial diseases. 136,137 In addition, feces and other excreta are a large source of bacterial survival in large numbers in humans and animals, and the use of ddPCR allows quantitative analysis of fecal–oral pathogens in human and animal feces. Some samples of human origin also include saliva and nasopharyngeal swab samples, and compared to qPCR, ddPCR has better reproducibility and better specificity, shorter detection time, and better sensitivity.…”