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Development and Clinical Validation of a Droplet Digital PCR Method for Detection of Acinetobacter baumannii and Klebsiella pneumonia in Patients with Suspected Bloodstream Infections
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  • Yang Zheng,
  • Jun Jin,
  • Ziqiang Shao,
  • Jingquan Liu,
  • Run Zhang,
  • Renhua Sun,
  • Bangchuan Hu
Yang Zheng
Zhejiang Provincial People's Hospital
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Jun Jin
Zhejiang Provincial People's Hospital
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Ziqiang Shao
Zhejiang Provincial People's Hospital
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Jingquan Liu
Zhejiang Provincial People's Hospital
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Run Zhang
Zhejiang Provincial People's Hospital
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Renhua Sun
Zhejiang Provincial People's Hospital
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Bangchuan Hu
Zhejiang Provincial People's Hospital
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Abstract

The relatively long turnaround time and low sensitivity of traditional blood culture may delay the effective antibiotic therapy in patients with bloodstream infection (BSI). To reduce the morbidity and mortality of BSI, a rapid and sensitive pathogen detection method is urgently required. Acinetobacter baumannii and Klebsiella pneumonia are two major microorganisms responsible for BSI. Here we reported a novel droplet digital PCR (ddPCR) method that can detect A. baumannii and K. pneumonia in whole blood samples within 4 h, with a specificity of 100% for each strain and limit of detection at 0.93 copies/microliter for A. baumannii and 0.27 copies/microliter for K. pneumonia. Clinical validation in 170 patients with suspected BSIs showed that, compared with blood culture that reported 4 (2.4%) A. baumannii cases and 7 (4.1%) K. pneumonia cases, ddPCR detected 23 (13.5%) A. baumannii cases, 26 (15.3%) K. pneumonia cases, and 4 (2.4%) dual infection cases, including the 11 positive patients reported by blood culture. In addition, the positive patients reported by ddPCR alone (n = 42) had significantly lower serum concentrations of procalcitonin and lactate, SOFA and APACHE II scores, and 28-day mortality than those reported by both blood culture and ddPCR (n = 11), suggesting that patients with less severe manifestations can potentially benefit from the guidance of ddPCR results. In conclusion, our study suggests that ddPCR represents a sensitive and rapid method to identify causal pathogens in blood samples and to guide the treatment decisions in the early stage of BSI.

Peer review status:UNDER REVIEW

06 May 2021Submitted to MicrobiologyOpen
07 May 2021Submission Checks Completed
07 May 2021Assigned to Editor
19 May 2021Reviewer(s) Assigned
09 Jun 2021Review(s) Completed, Editorial Evaluation Pending
11 Jun 2021Editorial Decision: Revise Major
26 Jun 20211st Revision Received
28 Jun 2021Submission Checks Completed
28 Jun 2021Assigned to Editor
28 Jun 2021Review(s) Completed, Editorial Evaluation Pending
28 Jun 2021Reviewer(s) Assigned