Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry

Abstract Early detection of drug resistance contributes to combating drug-resistant bacteria and improving patient outcomes. Microbial testing in the laboratory is essential for treating infectious diseases because it can provide critical information related to identifying pathogenic bacteria and th...

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Autores principales:	Takahiro Sawada, Masayuki Katayama, Shogo Takatani, Yoshiyuki Ohiro
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/679d7f754f414916b2ccd272b9746d49
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spelling	oai:doaj.org-article:679d7f754f414916b2ccd272b9746d492021-12-02T14:06:56ZEarly detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry10.1038/s41598-021-82186-42045-2322https://doaj.org/article/679d7f754f414916b2ccd272b9746d492021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82186-4https://doaj.org/toc/2045-2322Abstract Early detection of drug resistance contributes to combating drug-resistant bacteria and improving patient outcomes. Microbial testing in the laboratory is essential for treating infectious diseases because it can provide critical information related to identifying pathogenic bacteria and their resistance profiles. Despite these clinical requirements, conventional phenotypic testing is time-consuming. Additionally, recent rapid drug resistance tests are not compatible with fastidious bacteria such as Streptococcus and Haemophilus species. In this study, we validated the feasibility of direct bacteria counting using highly sensitive quantitative flow cytometry. Furthermore, by combining flow cytometry and a nucleic acid intercalator, we constructed a highly sensitive method for counting viable fastidious bacteria. These are inherently difficult to measure due to interfering substances from nutrients contained in the medium. Based on the conventional broth microdilution method, our method acquired a few microliter samples in a time series from the same microplate well to exclude the growth curve inconsistency between the samples. Fluorescent staining and flow cytometry measurements were completed within 10 min. Therefore, this approach enabled us to determine antimicrobial resistance for these bacteria within a few hours. Highly sensitive quantitative flow cytometry presents a novel avenue for conducting rapid antimicrobial susceptibility tests.Takahiro SawadaMasayuki KatayamaShogo TakataniYoshiyuki OhiroNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Takahiro Sawada Masayuki Katayama Shogo Takatani Yoshiyuki Ohiro Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry
description	Abstract Early detection of drug resistance contributes to combating drug-resistant bacteria and improving patient outcomes. Microbial testing in the laboratory is essential for treating infectious diseases because it can provide critical information related to identifying pathogenic bacteria and their resistance profiles. Despite these clinical requirements, conventional phenotypic testing is time-consuming. Additionally, recent rapid drug resistance tests are not compatible with fastidious bacteria such as Streptococcus and Haemophilus species. In this study, we validated the feasibility of direct bacteria counting using highly sensitive quantitative flow cytometry. Furthermore, by combining flow cytometry and a nucleic acid intercalator, we constructed a highly sensitive method for counting viable fastidious bacteria. These are inherently difficult to measure due to interfering substances from nutrients contained in the medium. Based on the conventional broth microdilution method, our method acquired a few microliter samples in a time series from the same microplate well to exclude the growth curve inconsistency between the samples. Fluorescent staining and flow cytometry measurements were completed within 10 min. Therefore, this approach enabled us to determine antimicrobial resistance for these bacteria within a few hours. Highly sensitive quantitative flow cytometry presents a novel avenue for conducting rapid antimicrobial susceptibility tests.
format	article
author	Takahiro Sawada Masayuki Katayama Shogo Takatani Yoshiyuki Ohiro
author_facet	Takahiro Sawada Masayuki Katayama Shogo Takatani Yoshiyuki Ohiro
author_sort	Takahiro Sawada
title	Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry
title_short	Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry
title_full	Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry
title_fullStr	Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry
title_full_unstemmed	Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry
title_sort	early detection of drug-resistant streptococcus pneumoniae and haemophilus influenzae by quantitative flow cytometry
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/679d7f754f414916b2ccd272b9746d49
work_keys_str_mv	AT takahirosawada earlydetectionofdrugresistantstreptococcuspneumoniaeandhaemophilusinfluenzaebyquantitativeflowcytometry AT masayukikatayama earlydetectionofdrugresistantstreptococcuspneumoniaeandhaemophilusinfluenzaebyquantitativeflowcytometry AT shogotakatani earlydetectionofdrugresistantstreptococcuspneumoniaeandhaemophilusinfluenzaebyquantitativeflowcytometry AT yoshiyukiohiro earlydetectionofdrugresistantstreptococcuspneumoniaeandhaemophilusinfluenzaebyquantitativeflowcytometry
_version_	1718391967043289088

Early detection of drug-resistant Streptococcus pneumoniae and Haemophilus influenzae by quantitative flow cytometry

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