Comparison of microarray and lysis filtration combined with MALDI-TOF MS methods for rapid diagnosis of gram-positive bacteria

Ayse Arslan; Mehmet Soylu; Mehmet Ali Özinel

doi:10.19161/etd.1642792

EN TR

Comparison of microarray and lysis filtration combined with MALDI-TOF MS methods for rapid diagnosis of gram-positive bacteria

Abstract

Aim: Early diagnosis of sepsis is crucial for patients’ prognosis. The primary objective of this study is to compare three methods for identifying bacteria in blood cultures: MALDI-TOF MS post-culture, MALDI-TOF MS after lysis filtration (LFM), and the Verigene System BC-GP microarray. Furthermore, the study evaluates the resistance profiles using the microarray method. Materials and Methods: A total of 40 blood culture bottles positive for Gram-positive bacteria were collected from adult patients in the intensive care unit (ICU) of a university hospital. After detecting positive signals, we applied three identification methods: MALDI-TOF MS from subcultures, MALDI-TOF MS after lysis filtration directly from the liquid sample, and the Verigene System BC-GP microarray (Nanosphere Inc. Northbrook, USA) test. Results: The consistency of LFM identification and the microarray method compared to post-culture MALDI-TOF MS was 75% (30/40) and 90% (36/40), respectively. The agreement between LFM and the microarray method was 83.3% (30/36). All three methods were consistent in 67.5% (27/40) of the bottles. Conclusion: The study indicates that both lysis filtration and microarray methods are effective for rapid diagnosis in sepsis patients. Each method has its own advantages and limitations. Lysis filtration combined with MALDI-TOF MS offers a cost-efficient, fast, and broad-database approach, providing results in approximately 9-10 hours instead of the conventional 48-72 hours when combined with automatic susceptibility testing.

Keywords

Gram-pozitif bakterilerin hızlı tanısında mikroarray ve lizis filtrasyon sonrası MALDI-TOF MS yöntemlerinin karşılaştırılması

Öz

Amaç: Sepsisin erken tanısı hastanın prognozu açısından çok önemlidir. Bu nedenle, etken mikroorganizmanın hızlı bir şekilde belirlenmesi ve uygun tedaviye başlanması kritik öneme sahiptir. Bu çalışmanın amacı, kan kültürlerinde üreyen Gram-pozitif bakterilerin tanımlanmasında kullanılan üç yöntemi karşılaştırmaktır: kültür sonrası MALDI-TOF MS tanımlama yöntemi, lizis filtrasyon sonrası MALDI-TOF MS (LFM) tanımlama yöntemi ve mikroarray yöntemi. Ayrıca, mikroarray yöntemi ile etkenlerin direnç profillerinin belirlenmesi hedeflenmiştir. Gereç ve Yöntem: Çalışmaya, bir üniversite hastanesinin erişkin yoğun bakım ünitelerinden alınan ve Gram-pozitif bakteri üremesi saptanan 40 kan kültürü şişesi dahil edilmiştir. Kan kültürü şişesinin pozitif sinyal vermesinden sonra, üç farklı tanımlama yöntemi uygulanmıştır: Subkültür yapıldıktan sonra kültür kolonisinden MALDI-TOF MS ile tanımlama, şişeden alınan örneğe lizis filtrasyon uygulandıktan sonra MALDI-TOF MS ile tanımlama ve Nanosphere Verigene System BC-GP mikroarray (Nanosphere Inc. Northbrook, USA) test yöntemi. Elde edilen sonuçlar karşılaştırılmıştır. Bulgular: Kültür sonrası MALDI TOF MS ile LFM tanımlaması arasındaki uyum oranı %75 (30/40) olarak hesaplanmıştır. Kültür sonrası MALDI TOF MS ile mikroarray sonuçları arasındaki uyum %90 (36/40) bulunmuştur. LFM ile mikroarray yöntemleri arasındaki benzerlik ise %83,3 (30/36) olarak belirlenmiştir. Üç yöntemin de tutarlı sonuç verdiği örneklerin oranı %67,5 (27/40) olarak hesaplanmıştır. Sonuç: Çalışma bulguları, sepsisli olgularda hem lizis filtrasyon hem de mikroarray test yöntemlerinin hızlı tanı açısından etkili olabileceğini ortaya koymuştur. Her iki yöntemin de avantajları ve sınırlılıkları mevcuttur. Lizis filtrasyon sonrası uygulanan MALDI-TOF MS yöntemi; düşük maliyeti, kısa sürede sonuç vermesi, geniş veri tabanı desteği ve Gram boyama gerektirmemesi gibi özellikleriyle, kütle spektrometrisi altyapısına sahip laboratuvarlar için uygun bir seçenek olarak değerlendirilmiştir. Ayrıca, lizis filtrasyon ile elde edilen süspansiyondan otomatik duyarlılık testlerinin yapılabilmesi sayesinde, yaklaşık 9–10 saat içinde tanı ve duyarlılık sonuçlarına ulaşılabileceği ve bu sürenin konvansiyonel yöntemlerle karşılaştırıldığında belirgin ölçüde daha kısa olduğu sonucuna varılmıştır.

Anahtar Kelimeler

Supporting Institution

Ege Üniversitesi Bilimsel Araştırmalar Etik Kurulu

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Details

Primary Language

English

Subjects

Medical Bacteriology

Journal Section

Research Article

Authors

Ayse Arslan ^*
0000-0002-4934-4889
Türkiye

Mehmet Soylu
0000-0002-9145-1506
Türkiye

Mehmet Ali Özinel
0000-0003-4142-4339
Türkiye

Publication Date

September 8, 2025

Submission Date

February 19, 2025

Acceptance Date

May 6, 2025

Published in Issue

Year 2025 Volume: 64 Number: 3

DOI

https://doi.org/10.19161/etd.1642792

IZ

https://izlik.org/JA49JU85HM

Cite

RIS / Bibtex

Vancouver

1.Ayse Arslan, Mehmet Soylu, Mehmet Ali Özinel. Comparison of microarray and lysis filtration combined with MALDI-TOF MS methods for rapid diagnosis of gram-positive bacteria. EJM. 2025 Sep. 1;64(3):460-7. doi:10.19161/etd.1642792