Rapid detection of Staphylococcus aureus in blood culture samples using human IgG-based lateral flow assay

Author:

Srisrattakarn Arpasiri1ORCID,Charoensri Nicha1ORCID,Prompipak Jeerati1ORCID,Ouancharee Wajeeorn1,Saiboonjan Bhanubong2ORCID,Tippayawat Patcharaporn1ORCID,Chanawong Aroonwadee1ORCID,Wonglakorn Lumyai3,Kanwattanee Ekgarak4,Piyapatthanakul Sirikan4,Masmalai Thitimar5,Ariyapim Anisara5,Kendal Rinjong Promson6,Lulitanond Aroonlug1ORCID

Affiliation:

1. Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand

2. Center for Innovation and Standard for Medical Technology and Physical Therapy, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand

3. Clinical Microbiology Unit, Srinagarind Hospital, Khon Kaen University, Khon Kaen, Thailand

4. Clinical Microbiology Laboratory, The Queen Sirikit National Institute of Child Health, Bangkok, Thailand

5. Clinical Laboratory, Queen Sirikit Heart Center of the Northeast, Khon Kaen University, Khon Kaen, Thailand

6. Khon Kaen Hospital, Khon Kaen, Thailand

Abstract

ABSTRACT Staphylococcus aureus is one of the most common pathogens. The conventional workflow for identifying this organism is time-consuming and takes up to several days. Therefore, we developed a colloidal gold-based lateral flow immunoassay (LFIA) using human IgG as a conjugated antibody to detect S. aureus . One hundred and thirty-eight clinical isolates, including 79 S . aureus and 59 non- S . aureus were spiked in blood samples, and incubated at 37°C for 24 h. The bacterial antigens were simply extracted before being tested by the developed LFIA strips. The results were read by the naked eye within 15 min. Conventional PCR was used as a reference method. The sensitivity and specificity of the developed LFIA were 100% (95% CI: 94.2%–100.0% and 92.4%–100.0%, respectively) in spiked blood culture samples. The detection limits of the LFIA for the purified protein A and bacterial colonies were 10 −3 µg/mL and 10 7 CFU/mL, respectively. The performance of the LFIA testing in 221 bacterial colony isolates and 118 positive blood culture bottles from three hospitals by their medical technologists showed 98.1% (95% CI: 94.1%–99.5%) and 89.7% (95% CI: 79.3%–95.4%) sensitivity, respectively. The LFIA is a quick, easy, and sensitive method for detecting S. aureus without expensive equipment. It might have the potential for early diagnosis of routine service in low-resource laboratories, leading to a rapid and effective treatment. IMPORTANCE In this study, we modified our previously developed lateral flow immunoassay (LFIA) test for the detection of Staphylococcus aureus by using an in-house human IgG as a conjugated antibody instead of the specific commercial antibody. It gave comparable results to the former developed-LFIA test and helped cost reduction.

Funder

Khon Kaen University

Publisher

American Society for Microbiology

Subject

Infectious Diseases,Cell Biology,Microbiology (medical),Genetics,General Immunology and Microbiology,Ecology,Physiology

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