ArticlePLoS neglected tropical diseases2023
Development and evaluation of a droplet digital PCR assay to detect Brucella in human whole blood.
Article in PLoS neglected tropical diseases, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers, 2 of them syntheses that pooled it.
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Who cites it
13 citing papers in PubMed, 2 syntheses or guidelines pooled it, 12 citations in OpenAlex.
- Epidemiology of Brucellosis Among Deer in China From 1978 to 2025: A Systematic Review and Meta-Analysis.Transboundary and emerging diseases · 2026Pooled it
- Research hotspots and development trends ofFrontiers in microbiology · 2025Pooled it
- Review
- Molecular Diagnostics of Infectious Diseases.Yonago acta medica · 2026Review
- Ultrasensitive Detection and Its Potential Applications in the Diagnosis of Brucellosis.Current microbiology · 2026Review
- The Evolution and Applications of Molecular Diagnostics in Veterinary and Clinical Medicine: From Traditional Methods to Emerging Technologies.Molecular biotechnology · 2026Review
- Prevalence and detection ofFrontiers in cellular and infection microbiology · 2026Article
- Large-scale evaluation of bacteriological-based method and qPCR performance for Brucellosis diagnosis in livestock using Bayesian latent class analysis.The veterinary quarterly · 2025Article
- Delayed diagnosis of pediatric brucellosis: a case study and literature review.Annals of medicine and surgery (2012) · 2025Article
- Specific detection of Brucella spp. from slaughter-aged livestock using a dual priming oligonucleotide system.BMC microbiology · 2025Article
- Detection and quantification ofThe veterinary quarterly · 2024Article
- Present and Future Applications of Digital PCR in Infectious Diseases Diagnosis.Diagnostics (Basel, Switzerland) · 2024Review
- Development of a duplex droplet digital PCR assay for the detection ofMicrobiology spectrum · 2024Article
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Authors and funding
10 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundWith the development of domestic animal husbandry, the spread of brucellosis has accelerated, and the scope of the epidemic has expanded. The timely and accurate diagnosis of human brucellosis continues to challenge clinicians in endemic areas. Droplet digital PCR (ddPCR) technology can quickly and accurately determine DNA load in samples, providing laboratory evidence for diagnosis, prognosis and management of brucellosis patients. In this study, a ddPCR method was established to accurately quantify Brucella DNA load in whole blood samples, and its diagnostic, prognostic, and therapeutic value for human brucellosis was evaluated.
methodsAnnealing temperature, primers, and probe targeting the Brucella bcsp31 gene were optimised, and the sensitivity, specificity and repeatability of the ddPCR assay were assessed using 94 whole blood samples from 61 confirmed and 33 suspected cases. Results were compared with those of quantitative PCR (qPCR). Nine follow-up brucellosis patients were also analysed by the two methods after 2 and 6 months of treatment.
resultsOptimal primer and probe concentrations were 800 nmol/L and 400 nmol/L, respectively, and the optimal annealing temperature was 55.3 °C. The ddPCR results showed that the limit of detection was 1.87 copies per reaction, with high repeatability. The positive rates for ddPCR and qPCR were 88.5% and 75.4% among 61 serum agglutination test (SAT) positive patients. In addition, 57.6% (19/33) of suspected sero-negative samples were positive by ddPCR, but only 36.3% (12/33) were positive by qPCR. Analysis of nine post-therapy follow-up brucellosis patients revealed that the Brucella DNA load in the whole blood samples decreased after 2 and 6 months of treatment, and was slightly increased following relapse and continuous exposure.
conclusionThe ddPCR assay showed good accuracy for whole blood samples, and could be a potential diagnostic and prognostic tool for detecting Brucella.
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