We use cookies to understand how you use our site and to improve your experience. This includes personalizing content and advertising. To learn more, click here. By continuing to use our site, you accept our use of cookies. Cookie Policy.

Features Partner Sites Information LinkXpress
Sign In
Advertise with Us
PURITAN MEDICAL

Download Mobile App




Events

09 Apr 2024 - 12 Apr 2024
15 Apr 2024 - 17 Apr 2024
23 Apr 2024 - 26 Apr 2024

MALDI-TOF MS Identifies Oomycete Causing Pythiosis

By LabMedica International staff writers
Posted on 12 Dec 2018
Print article
Image: The UltrafleXtreme MALDI-TOF/TOF mass spectrometer (Photo courtesy of Bruker Daltonics).
Image: The UltrafleXtreme MALDI-TOF/TOF mass spectrometer (Photo courtesy of Bruker Daltonics).
Pythiosis is an invasive, difficult-to-treat, life-threatening infectious disease caused by Pythium insidiosum, a member of the unique group of fungus-like microorganisms called oomycetes. The disease has been increasingly reported worldwide.

In the past decade, the matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has emerged as a novel and powerful diagnostic tool for facilitating the clinical identification of many pathogenic microorganisms, including bacteria and fungi.

Scientists at the Mahidol University (Bangkok, Thailand) isolated a total of 13 strains of P. insidiosum, isolated from eight humans and five animals with pythiosis, from different geographic locations. All organisms were maintained on Sabouraud dextrose agar at 25 °C. Several small portions of a colony of each organism were transferred to a 50-mL flask containing 10 mL Sabouraud dextrose broth, and incubated at 37 °C for one week, before harvesting fungal material for protein extraction.

Protein was extracted from harvested organisms and was spotted onto a clean ground steel target plate in 40 replicates (for generating a MALDI-TOF MS database of P. insidiosum) or five replicates (for assessing the MALDI-TOF MS for identification of P. insidiosum), air dried at room temperature before being processed. After the matrix solution was air dried at room temperature, the sample was promptly analyzed, using a Bruker ultrafleXtreme mass spectrometer. Genomic DNA (gDNA) templates were extracted from the organisms and subjected to single nucleotide polymorphism-based multiplex polymerase chain reaction (PCR).

The team reported that the MALDI-TOF MS accurately identified all 13 P. insidiosum strains tested, at the species level. Mass spectra of P. insidiosum did not match any other microorganisms, including fungi (i.e., Aspergillus species, Fusarium species, and fungal species of the class Zygomycetes), which have similar microscopic morphologies with this oomycete. MALDI-TOF MS- and rDNA sequence-based biotyping methods consistently classified P. insidiosum into three groups: Clade-I (American strains), II (Asian and Australian strains), and III (mostly Thai strains).

The authors concluded that MALDI-TOF MS has been successfully used for identification and biotyping of P. insidiosum. The obtained mass spectral database allows clinical microbiology laboratories, well equipped with a MALDI-TOF mass spectrometer, to conveniently identify P. insidiosum, without requiring any pathogen-specific reagents (i.e., antigen, antibody or primers). The study was published in the December 2018 issue of the International Journal of Infectious Diseases.

Related Links:
Mahidol University

Platinum Member
COVID-19 Rapid Test
OSOM COVID-19 Antigen Rapid Test
One Step HbA1c Measuring System
GREENCARE A1c
POCT Fluorescent Immunoassay Analyzer
FIA Go
Gold Member
Real-time PCR System
GentierX3 Series

Print article

Channels

Clinical Chemistry

view channel
Image: Reaching speeds up to 6,000 RPM, this centrifuge forms the basis for a new type of inexpensive, POC biomedical test (Photo courtesy of Duke University)

POC Biomedical Test Spins Water Droplet Using Sound Waves for Cancer Detection

Exosomes, tiny cellular bioparticles carrying a specific set of proteins, lipids, and genetic materials, play a crucial role in cell communication and hold promise for non-invasive diagnostics.... Read more

Molecular Diagnostics

view channel
Image: MOF materials efficiently enrich cfDNA and cfRNA in blood through simple operational process (Photo courtesy of Science China Press)

Blood Circulating Nucleic Acid Enrichment Technique Enables Non-Invasive Liver Cancer Diagnosis

The ability to diagnose diseases early can significantly enhance the effectiveness of clinical treatments and improve survival rates. One promising approach for non-invasive early diagnosis is the use... Read more

Hematology

view channel
Image: The low-cost portable device rapidly identifies chemotherapy patients at risk of sepsis (Photo courtesy of 52North Health)

POC Finger-Prick Blood Test Determines Risk of Neutropenic Sepsis in Patients Undergoing Chemotherapy

Neutropenia, a decrease in neutrophils (a type of white blood cell crucial for fighting infections), is a frequent side effect of certain cancer treatments. This condition elevates the risk of infections,... Read more

Pathology

view channel
Image: The OvaCis Rapid Test discriminates benign from malignant epithelial ovarian cysts (Photo courtesy of INEX)

Intra-Operative POC Device Distinguishes Between Benign and Malignant Ovarian Cysts within 15 Minutes

Ovarian cysts represent a significant health issue for women globally, with up to 10% experiencing this condition at some point in their lives. These cysts form when fluid collects within a thin membrane... Read more
Copyright © 2000-2024 Globetech Media. All rights reserved.