Let them fly or light them up: matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry and fluorescence in situ hybridization (FISH)

• Published in: APMIS : acta pathologica, microbiologica et immunologica Scandinavica microbiologica et immunologica Scandinavica, 2004-11, Vol.112 (11-12), p.856-885
• Main Authors: SCHWEICKERT, BIRGITTA, MOTER, ANNETTE, LEFMANN, MICHAEL, GÖBEL, ULF B.
• Format: Article
• Language: English
• Subjects: 16S rRNA; Bacteria - genetics; Bacterial Infections - diagnosis; Bacterial Infections - microbiology; Bacteriological Techniques - methods; clinical microbiology; FISH; Gastrointestinal Tract - microbiology; Genome, Bacterial; Humans; In Situ Hybridization, Fluorescence - methods; MALDI-TOF; Mouth - microbiology; Respiratory System - microbiology; RNA, Bacterial - chemistry; RNA, Bacterial - genetics; RNA, Ribosomal, 16S - genetics; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
• ISSN: 0903-4641; 1600-0463
• DOI: 10.1111/j.1600-0463.2004.apm11211-1210.x

This review focuses on clinical bacteriology and by and large does not cover the detection of fungi, viruses or parasites. It discusses two completely different but complementary approaches that may either supplement or replace classic culture‐based bacteriology. The latter view may appear provocative in the light of the actual market penetration of molecular genetic testing in clinical bacteriology. Despite its elegance, high specificity and sensitivity, molecular genetic diagnostics has not yet reached the majority of clinical laboratories. The reasons for this are manifold: Many microbiologists and medical technologists are more familiar with classical microbiological methods than with molecular biology techniques. Culture‐based methods still represent the work horse of everyday routine. The number of available FDA‐approved molecular genetic tests is limited and external quality control is still under development. Finally, it appears difficult to incorporate genetic testing in the routine laboratory setting due to the limited number of samples received or the lack of appropriate resources. However, financial and time constraints, particularly in hospitals as a consequence of budget cuts and reduced length of stay, lead to a demand for significantly shorter turnaround times that cannot be met by culture‐dependent diagnosis. As a consequence, smaller laboratories that do not have the technical and personal equipment required for molecular genetic amplification techniques may adopt alternative methods such as fluorescence in situ hybridization (FISH) that combines easy‐to‐perform molecular hybridization with microscopy, a technique familiar to every microbiologist. FISH is hence one of the technologies presented here. For large hospital or reference laboratories with a high sample volume requiring massive parallel high‐throughput testing we discuss matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF) of nucleic acids, a technology that has evolved from the post‐genome sequencing era, for high‐throughput sequence variation analysis (1, 2).