Usefulness of the MALDI-TOF MS technology with membrane filter protocol for the rapid identification of microorganisms in perioperative drainage fluids of hepatobiliary pancreatic surgery

Kazuyuki Sogawa; Shigetsugu Takano; Takayuki Ishige; Hideyuki Yoshitomi; Shingo Kagawa; Katsunori Furukawa; Tsukasa Takayashiki; Satoshi Kuboki; Fumio Nomura; Masayuki Ohtsuka

doi:10.1371/journal.pone.0246002

Usefulness of the MALDI-TOF MS technology with membrane filter protocol for the rapid identification of microorganisms in perioperative drainage fluids of hepatobiliary pancreatic surgery

PLoS One. 2021 Feb 4;16(2):e0246002. doi: 10.1371/journal.pone.0246002. eCollection 2021.

Affiliations

¹ Department of Biochemistry, School of Life and Environmental Science, Azabu University, Kanagawa, Japan.
² Department of General Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan.
³ Division of Laboratory Medicine, Chiba University Hospital, Chiba, Japan.
⁴ Divisions of Clinical Mass Spectrometry and Clinical Genetics, Chiba University Hospital, Chiba, Japan.

Abstract

Surgical site infections (SSIs) are significant and frequent perioperative complications, occurring due to the contamination of the surgical site. The late detection of SSIs, especially organ/space SSIs which are the more difficult to treat, often leads to severe complications. An effective method that can identify bacteria with a high accuracy, leading to the early detection of organ/space SSIs, is needed. Ninety-eight drainage fluid samples obtained from 22 patients with hepatobiliary pancreatic disease were analyzed to identify microorganisms using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) with a new membrane filtration protocol and rapid BACpro® pretreatment compared to sole rapid BACpro® pretreatment. The levels of detail of rapid BACpro® pretreatment with or without filtration were also evaluated for the accuracy of bacterial identification. We found that reliable scores for E. coli and E. faecalis were obtained by inoculation with 1.0 × 104 CFU/ml after preparation of the membrane filter with rapid BACpro®, indicating approximately 10-folds more sensitive compared to sole rapid BACpro® pretreatment in drainage fluid specimens. Among 60 bacterial positive colonies in drainage fluid specimens, the MALDI-TOF MS and the membrane filtration with rapid BACpro® identified 53 isolates (88.3%) with a significantly higher accuracy, compared to 25 isolates in the rapid BACpro® pretreatment group (41.7%) (p < 0.001). Among the 78 strains, 14 enteric Gram-negative bacteria (93.0%) and 55 Gram-positive cocci (87.3%) were correctly identified by the membrane filtration with rapid BACpro® with a high reliability. This novel protocol could identify bacterial species within 30 min, at $2-$3 per sample, thus leading to cost and time savings. MALDI-TOF MS with membrane filter and rapid BACpro® is a quick and reliable method for bacterial identification in drainage fluids. The shortened analysis time will enable earlier selection of suitable antibiotics for treatment of organ/space SSIs to improve patients' outcomes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aged
Animals
Bacterial Typing Techniques / methods*
Drainage*
Female
Filtration*
Hepatopancreas / surgery*
Humans
Male
Membranes, Artificial*
Perioperative Period*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization*
Time Factors

Substances

Membranes, Artificial

Grants and funding

This work was supported by a Grant-in-Aid for Scientific Research (KAKENHI): KIBAN C (grant no. 19K07947 to KS, grant no. 19K09113 to MO, ST, SaK), and KIBAN B (grant no. 19H03725 to ST, ShK, and MO). The URL of Grant-in-Aid for Scientific Research (KAKENHI) is as follows; https://www.jsps.go.jp/english/e-grants/ The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.