Scientists from Necker-Enfants Malades Hospital (AP-HP), the Institut Pasteur, Université Paris Cité, Inserm, Université Paris-Est Créteil and the Alfort National Veterinary School, coordinated by Professor Marc Eloit and Dr. Anne Jamet in collaboration with Dr. Jacques Fourgeaud and Beatrice Regnault, studied the role of global genetic characterization of samples (or non-targeted metagenomics) using next-generation sequencing (mNGS) in diagnosing infections. The results of the study were published on December 1, 2023 in the journal Lancet Microbe.
The identification of microorganisms involved in infection is a major challenge in ensuring that patients receive optimal treatment. Recent epidemics have shown the importance of a tool capable of detecting new or unexpected pathogens and those with rapidly evolving genomes. Currently, the search for infectious agents is mostly ‘targeted’ and requires prior knowledge of the possible causes of infection. In many cases however, no microorganism is identified by first-line testing and the cause of the infection remains unknown, leading to suboptimal treatment.
Metagenomics1 using next-generation sequencing (mNGS) can identify a wide range of pathogens, including rare or novel microorganisms. This study aims to improve use of this innovative microbial identification technique, which remains complex and costly for hospital laboratories as it requires cross-disciplinary skills ranging from specific sample preparation to bioinformatics analysis of a large number of sequences. The hospital team drew on the technical and IT skills of the Institut Pasteur’s Pathogen Discovery Laboratory2 and worked to make mNGS available within the AP-HP and to practitioners in healthcare centers in mainland France and overseas.
In this study, 742 samples were collected for mNGS analysis from 523 patients between October 29, 2019 and November 7, 2022. Samples were accompanied by a mandatory prescription form completed by the physician, indicating the level of clinical suspicion of infection.
The results relate to a panel whose initial suspicion of infection was either high (63%) or low (37%). In 117 patient samples where infection was strongly suspected – i.e. 25% of samples where infection was strongly suspected according to the practitioners’ preliminary assessment – causative or potentially causative pathogens were detected.
The diagnostic yield of mNGS was particularly high in immunocompromised patients and in patients with neurological disorders where brain biopsies were available. In fact, mNGS more easily detects a causative or potentially causative pathogenic virus in brain biopsies than in cerebrospinal fluid, which is traditionally used because it is easier to obtain. Furthermore, the study showed that stool analyses could be used to investigate not only digestive disorders, but also hepatitis and various neurological symptoms.
In addition, the clinical performance of mNGS compares favorably to conventional microbiology. Together with future studies, the results of this prospective observational study will help to define the role of mNGS in decision making relating to diagnosis and treatment.
“The Microbiology Laboratory (Prof. Leruez-Ville), in conjunction with the Department of Infectious and Tropical Diseases (Prof. Lortholary and Prof. Lecuit) and the Pediatric Immunohematology and Rheumatology Department (Prof. Quartier dit Maire and Prof. Neven) at Necker-Enfants Malades Hospital (AP-HP), has developed extensive expertise in the management of immunocompromised patients prone to infections caused by unusual microorganisms. The Institut Pasteur’s Pathogen Discovery Laboratory2, which was led by Prof. Marc Eloit at the time of the study, had already optimized the use of high-throughput sequencing for pathogen discovery by developing the technique for both sample preparation methods and bioinformatics analysis tools. This collaboration between Necker-Enfants Malades (AP-HP) and the Institut Pasteur was therefore well placed to identify novel causes of infection in cases where conventional techniques were missing diagnoses,” explains Dr. Anne Jamet, the study’s final author, who is head of mNGS at Necker-Enfants Malades Hospital (AP-HP) and a researcher at the Institut Necker-Enfants Malades (AP-HP).
“Our instincts have translated into reliable diagnoses in everyday practice as well as several noteworthy discoveries, including the identification of a new virus responsible for hepatitis,” adds Dr. Jacques Fourgeaud, first author of the study and lead virologist for mNGS at Necker-Enfants Malades Hospital (AP-HP).
“This sequencing-based tool is now indispensable for diagnosing patients with a suspected infection. We are now using it earlier and earlier in severe cases, particularly those involving the brain, and in immunocompromised adults and children,” says Prof. Olivier Lortholary, an infectious diseases specialist who is head of the Department of Infectious and Tropical Diseases at Necker-Enfants Malades Hospital (AP-HP) and co-author of the study.
“We're delighted to be able to contribute to better medical care, while at the same time increasing our knowledge of infectious diseases,” adds Prof. Marc Eloit, co-last author of the study, who led the Pathogen Discovery Laboratory2 until September 2023 and is now a visiting researcher at the Institut Pasteur. The study also provides an insight into the future of infection diagnostics. “The microorganisms we identify with the Necker microbiology laboratory will enable us to develop new tools to make sequencing technology faster and more accessible for front-line microbiological analysis while enhancing its potential for the discovery of new human pathogens,” says co-author Philippe Pérot, a research engineer in the Institut Pasteur's Pathogen Discovery Laboratory2 and lead for mNGS at the Institut Pasteur.
The study was funded by Necker-Enfants Malades Hospital (AP-HP) and the Institut Pasteur.
- Study of all genomes from a single environment and interactions between them
- The Institut Pasteur's Pathogen Discovery Laboratory was led by Prof. Marc Eloit until August 31, 2023. Dr. Nolwenn Dheilly took over as head of the laboratory on September 1, 2023
Source
Performance of clinical metagenomics in France: a prospective observational study, The Lancet Microbe, 1er décembre 2023
Jacques Fourgeaud, PharmD a,b, Béatrice Regnault, MSc *c,d, Vichita Ok, PharmD b, Nicolas Da Rocha, MSc c, Émilie Sitterlé, PharmD b, Meryem Mekouar, MD b, Hélène Faury, MD b, Catherine Milliancourt-Seels, BSc b, Florence Jagorel, BSc c, Delphine Chrétien, BSc c, Thomas Bigot, PhD d, Éric Troadec, BSc c, Isabelle Marques, BSc b, Alexandra Serris, MD e, Prof Danielle Seilhean, MD f,o, Prof Bénédicte Neven, MD g,h, Prof Pierre Frange, MD a,b, Agnès Ferroni, PharmD b, Prof Marc Lecuit, MD e,i,j, Prof Xavier Nassif, MD k, Prof Olivier Lortholary, MD † e,l, Marianne Leruez-Ville, MD † a,b, Philippe Pérot, PhD † c,l, Prof Marc Eloit, PhD ‡ c,m,n, Anne Jamet, MD ‡ b,k
a Université Paris Cité, FETUS, Paris, France
b Microbiology Department, AP-HP, Hôpital Necker, Paris, France
c Institut Pasteur, Université Paris Cité, Pathogen Discovery Laboratory, Paris, France
d Bioinformatics and Biostatistics Hub, Computational Biology Department, Institut Pasteur, Paris, France
e Université Paris Cité, Centre d'Infectiologie Necker-Pasteur, IHU Imagine, Hôpital Necker, Paris, France
f Département de Neuropathologie Raymond Escourolle, AP-HP-Sorbonne, Groupe Hospitalier Pitié-Salpêtrière, Paris, France
g Pediatric Hematology Immunology and Rheumatology Unit, AP-HP, Hôpital Necker, Paris, France
h Université Paris Cité, INSERM, Institut Imagine, Laboratory of Immunogenetics of Pediatric Autoimmune Diseases, Paris, France
i Institut Pasteur, Université de Paris, INSERM U1117, Biology of Infection Unit, Paris, France
j Institut Pasteur, National Reference Center and WHO Collaborating Center Listeria, Paris, France
k Université Paris Cité, CNRS, INSERM, Institut Necker-Enfants Malades, Team Pathogenesis of Systemic Infection, Paris, France
l Institut Pasteur, Centre National de Référence Mycoses Invasives et Antifongiques, Département de Mycologie, Labex IBEID, Paris, France
m Institut Pasteur, Université Paris Cité, The WOAH Collaborating Center for the Detection and Identification in Humans of Emerging Animal Pathogens, Paris, France
n École Nationale Vétérinaire d'Alfort, Université Paris-Est, Maisons-Alfort, France
o Institut du Cerveau–Paris Brain Institute–ICM, INSERM U1127, CNRS UMR7225, AP-HP, Sorbonne University, Pitié-Salpêtrière Hospital, Paris, France
