Affinity molecular assay for detecting Candida albicans using chitin affinity and RPA-CRISPR/Cas12a.

Candida albicans / genetics Humans Chitin / metabolism CRISPR-Cas Systems DNA, Fungal / genetics Sensitivity and Specificity Bronchoalveolar Lavage Fluid / microbiology Endodeoxyribonucleases / metabolism Candidiasis / diagnosis Bacterial Proteins CRISPR-Associated Proteins

Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
28 Oct 2024
Historique:
received: 21 02 2024
accepted: 18 10 2024
medline: 29 10 2024
pubmed: 29 10 2024
entrez: 29 10 2024
Statut: epublish

Résumé

Invasive fungal infections (IFIs) pose a significant threat to immunocompromised individuals, leading to considerable morbidity and mortality. Prompt and accurate diagnosis is essential for effective treatment. Here we develop a rapid molecular diagnostic method that involves three steps: fungal enrichment using affinity-magnetic separation (AMS), genomic DNA extraction with silicon hydroxyl magnetic beads, and detection through a one-pot system. This method, optimized to detect 30 CFU/mL of C. albicans in blood and bronchoalveolar lavage (BAL) samples within 2.5 h, is approximately 100 times more sensitive than microscopy-based staining. Initial validation using clinical samples showed 93.93% sensitivity, 100% specificity, and high predictive values, while simulated tests demonstrated 95% sensitivity and 100% specificity. This cost-effective, highly sensitive technique offers potential for use in resource-limited clinical settings and can be easily adapted to differentiate between fungal species and detect drug resistance.

Identifiants

DOI: 10.1038/s41467-024-53693-5 PMID: 39468064
pubmed: 39468064
doi: 10.1038/s41467-024-53693-5
pii: 10.1038/s41467-024-53693-5
doi:

Substances chimiques

Chitin 1398-61-4
DNA, Fungal 0
Cas12a protein EC 3.1.-
Endodeoxyribonucleases EC 3.1.-
Bacterial Proteins 0
CRISPR-Associated Proteins 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

9304

Subventions

Organisme : National Natural Science Foundation of China (National Science Foundation of China)
ID : 82072349

Informations de copyright

© 2024. The Author(s).

Références

Kainz, K., Bauer, M. A., Madeo, F. & Carmona-Gutierrez, D. Fungal infections in humans: the silent crisis. Micro Cell 7, 143–145 (2020).
doi: 10.15698/mic2020.06.718
Fang, W. et al. Diagnosis of invasive fungal infections: challenges and recent developments. J. Biomed. Sci. 30, 42 (2023).
pubmed: 37337179 pmcid: 10278348 doi: 10.1186/s12929-023-00926-2
Arvanitis, M., Anagnostou, T., Fuchs, B. B., Caliendo, A. M. & Mylonakis, E. Molecular and nonmolecular diagnostic methods for invasive fungal infections. Clin. Microbiol. Rev. 27, 490–526 (2014).
pubmed: 24982319 pmcid: 4135902 doi: 10.1128/CMR.00091-13
Alves, J., Alonso-Tarrés, C. & Rello, J. How to identify invasive candidemia in ICU—a narrative review. Antibiotics 11, 1804 (2022).
Lass-Flörl, C., Samardzic, E. & Knoll, M. Serology anno 2021-fungal infections: from invasive to chronic. Clin. Microbiol. Infect. 27, 1230–1241 (2021).
pubmed: 33601011 doi: 10.1016/j.cmi.2021.02.005
Wen, X., Chen, Q., Yin, H., Wu, S. & Wang, X. Rapid identification of clinical common invasive fungi via a multi-channel real-time fluorescent polymerase chain reaction melting curve analysis. Medicine 99, e19194 (2020).
pubmed: 32049856 pmcid: 7035122 doi: 10.1097/MD.0000000000019194
Scorzoni, L. et al. Antifungal therapy: new advances in the understanding and treatment of mycosis. Front. Microbiol. 8, 36 (2017).
pubmed: 28167935 pmcid: 5253656 doi: 10.3389/fmicb.2017.00036
Janbon, G., Quintin, J., Lanternier, F. & d’Enfert, C. Studying fungal pathogens of humans and fungal infections: fungal diversity and diversity of approaches. Genes Immun. 20, 403–414 (2019).
pubmed: 31019254 doi: 10.1038/s41435-019-0071-2
Wiederhold, N. P. Emerging fungal infections: new species, new names, and antifungal resistance. Clin. Chem. 68, 83–90 (2021).
pubmed: 34969112 pmcid: 9383166 doi: 10.1093/clinchem/hvab217
Lockhart, S. R. & Guarner, J. Emerging and reemerging fungal infections. Semin Diagn. Pathol. 36, 177–181 (2019).
pubmed: 31010605 doi: 10.1053/j.semdp.2019.04.010
Lockhart, S. R., Chowdhary, A. & Gold, J. A. W. The rapid emergence of antifungal-resistant human-pathogenic fungi. Nat. Rev. Microbiol. 21, 818–832 (2023).
pubmed: 37648790 doi: 10.1038/s41579-023-00960-9
Lenardon, M. D., Munro, C. A. & Gow, N. A. R. Chitin synthesis and fungal pathogenesis. Curr. Opin. Microbiol. 13, 416–423 (2010).
pubmed: 20561815 pmcid: 2923753 doi: 10.1016/j.mib.2010.05.002
Gow, N. A. R., Latge, J.-P. & Munro, C. A. The fungal cell wall: structure, biosynthesis, and function. Microbiol. Spectr. https://doi.org/10.1128/microbiolspec.FUNK-0035-2016 (2017).
Bonhomme, J. & d’Enfert, C. Candida albicans biofilms: building a heterogeneous, drug-tolerant environment. Curr. Opin. Microbiol. 16, 398–403 (2013).
pubmed: 23566895 doi: 10.1016/j.mib.2013.03.007
Steinfeld, L., Vafaei, A., Rösner, J. & Merzendorfer, H. Chitin prevalence and function in bacteria, fungi and protists. Adv. Exp. Med. Biol. 1142, 19–59 (2019).
pubmed: 31102241 doi: 10.1007/978-981-13-7318-3_3
Atkinson, E. M. & Long, S. R. Homology of Rhizobium meliloti NodC to polysaccharide polymerizing enzymes. Mol. Plant Microbe Interact. 5, 439–442 (1992).
pubmed: 1472720 doi: 10.1094/MPMI-5-439
Ren, Z. et al. Structural basis for inhibition and regulation of a chitin synthase from Candida albicans. Nat. Struct. Mol. Biol. 29, 653–664 (2022).
pubmed: 35788183 pmcid: 9359617 doi: 10.1038/s41594-022-00791-x
Bowman, S. M. & Free, S. J. The structure and synthesis of the fungal cell wall. Bioessays 28, 799–808 (2006).
pubmed: 16927300 doi: 10.1002/bies.20441
Haydour, Q. et al. Diagnosis of fungal infections. A systematic review and meta-analysis supporting American Thoracic Society practice guideline. Ann. Am. Thorac. Soc. 16, 1179–1188 (2019).
pubmed: 31219341 doi: 10.1513/AnnalsATS.201811-766OC
Mendonca, A., Santos, H., Franco-Duarte, R. & Sampaio, P. Fungal infections diagnosis—past, present and future. Res. Microbiol. 173, 103915 (2022).
pubmed: 34863883 doi: 10.1016/j.resmic.2021.103915
Kontoyiannis, D. P. Antifungal resistance: an emerging reality and a global challenge. J. Infect. Dis. 216, S431–S435 (2017).
pubmed: 28911044 doi: 10.1093/infdis/jix179
Ganguli, A. et al. A culture-free biphasic approach for sensitive and rapid detection of pathogens in dried whole-blood matrix. Proc. Natl. Acad. Sci. USA 119, e2209607119 (2022).
pubmed: 36161889 pmcid: 9546527 doi: 10.1073/pnas.2209607119
Katevatis, C., Fan, A. & Klapperich, C. M. Low concentration DNA extraction and recovery using a silica solid phase. PLoS One 12, e0176848 (2017).
pubmed: 28475611 pmcid: 5419563 doi: 10.1371/journal.pone.0176848
Kasahara, K., Ishikawa, H., Sato, S., Shimakawa, Y. & Watanabe, K. Development of multiplex loop-mediated isothermal amplification assays to detect medically important yeasts in dairy products. FEMS Microbiol. Lett. 357, 208–216 (2014).
pubmed: 24965944
Kourkoumpetis, T. K., Fuchs, B. B., Coleman, J. J., Desalermos, A. & Mylonakis, E. Polymerase chain reaction-based assays for the diagnosis of invasive fungal infections. Clin. Infect. Dis. 54, 1322–1331 (2012).
pubmed: 22362884 pmcid: 3491854 doi: 10.1093/cid/cis132
Zhao, Y., Chen, F., Li, Q., Wang, L. & Fan, C. Isothermal amplification of nucleic acids. Chem. Rev. 115, 12491–12545 (2015).
pubmed: 26551336 doi: 10.1021/acs.chemrev.5b00428
Li, J. & Macdonald, J. Advances in isothermal amplification: novel strategies inspired by biological processes. Biosens. Bioelectron. 64, 196–211 (2015).
pubmed: 25218104 doi: 10.1016/j.bios.2014.08.069
Lobato, I. M. & O’Sullivan, C. K. Recombinase polymerase amplification: basics, applications and recent advances. Trends Anal. Chem. 98, 19–35 (2018).
doi: 10.1016/j.trac.2017.10.015
Najjar, D. et al. A lab-on-a-chip for the concurrent electrochemical detection of SARS-CoV-2 RNA and anti-SARS-CoV-2 antibodies in saliva and plasma. Nat. Biomed. Eng. 6, 968–978 (2022).
pubmed: 35941191 pmcid: 9361916 doi: 10.1038/s41551-022-00919-w
Qin, C. et al. One-pot visual detection of African swine fever virus using CRISPR-Cas12a. Front Vet. Sci. 9, 962438 (2022).
pubmed: 35923823 pmcid: 9339671 doi: 10.3389/fvets.2022.962438
Kaminski, M. M., Abudayyeh, O. O., Gootenberg, J. S., Zhang, F. & Collins, J. J. CRISPR-based diagnostics. Nat. Biomed. Eng. 5, 643–656 (2021).
pubmed: 34272525 doi: 10.1038/s41551-021-00760-7
Chen, J. S. et al. CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science 360, 436–439 (2018).
pubmed: 29449511 pmcid: 6628903 doi: 10.1126/science.aar6245
Lu, S. et al. Fast and sensitive detection of SARS-CoV-2 RNA using suboptimal protospacer adjacent motifs for Cas12a. Nat. Biomed. Eng. 6, 286–297 (2022).
pubmed: 35314803 doi: 10.1038/s41551-022-00861-x
Arizti-Sanz, J. et al. Streamlined inactivation, amplification, and Cas13-based detection of SARS-CoV-2. Nat. Commun. 11, 5921 (2020).
pubmed: 33219225 pmcid: 7680145 doi: 10.1038/s41467-020-19097-x
Malcı, K., Walls, L. E. & Rios-Solis, L. Rational design of CRISPR/Cas12a-RPA based one-pot COVID-19 detection with design of experiments. ACS Synth. Biol. 11, 1555–1567 (2022).
pubmed: 35363475 pmcid: 9016756 doi: 10.1021/acssynbio.1c00617
Zeng, Q. et al. Rapid and sensitive Cas12a-based one-step nucleic acid detection with ssDNA-modified crRNA. Anal. Chim. Acta 1276, 341622 (2023).
pubmed: 37573099 doi: 10.1016/j.aca.2023.341622
Mehmood, M. A., Xiao, X., Hafeez, F. Y., Gai, Y. & Wang, F. Molecular characterization of the modular chitin binding protein Cbp50 from Bacillus thuringiensis serovar konkukian. Antonie Van. Leeuwenhoek 100, 445–453 (2011).
pubmed: 21647612 doi: 10.1007/s10482-011-9601-2
Barboza-Corona, J. E. et al. Cloning, sequencing, and expression of the chitinase gene chiA74 from Bacillus thuringiensis. Appl Environ. Microbiol 69, 1023–1029 (2003).
pubmed: 12571025 pmcid: 143672 doi: 10.1128/AEM.69.2.1023-1029.2003
Bormann, C. et al. Characterization of a novel, antifungal, chitin-binding protein from Streptomyces tendae Tü901 that interferes with growth polarity. J. Bacteriol. 181, 7421–7429 (1999).
pubmed: 10601197 pmcid: 94197 doi: 10.1128/JB.181.24.7421-7429.1999
Vaaje-Kolstad, G., Houston, D. R., Riemen, A. H. K., Eijsink, V. G. H. & van Aalten, D. M. F. Crystal structure and binding properties of the Serratia marcescens chitin-binding protein CBP21. J. Biol. Chem. 280, 11313–11319 (2005).
pubmed: 15590674 doi: 10.1074/jbc.M407175200
Muller, D. A., Corrie, S. R., Coffey, J., Young, P. R. & Kendall, M. A. Surface modified microprojection arrays for the selective extraction of the dengue virus NS1 protein as a marker for disease. Anal. Chem. 84, 3262–3268 (2012).
pubmed: 22424552 doi: 10.1021/ac2034387
Yang, Q. et al. A rapid, visible, and highly sensitive method for recognizing and distinguishing invasive fungal infections via CCP-FRET technology. ACS Infect. Dis. 7, 2816–2825 (2021).
pubmed: 34585580 doi: 10.1021/acsinfecdis.1c00393
Louie, A. et al. Pharmacodynamics of fluconazole in a murine model of systemic candidiasis. Antimicrob. Agents Chemother. 42, 1105–1109 (1998).
pubmed: 9593135 pmcid: 105753 doi: 10.1128/AAC.42.5.1105
Clancy, C. J. & Nguyen, M. H. Finding the “missing 50%“ of invasive candidiasis: how nonculture diagnostics will improve understanding of disease spectrum and transform patient care. Clin. Infect. Dis. 56, 1284–1292 (2013).
pubmed: 23315320 doi: 10.1093/cid/cit006
Robbins, N., Caplan, T. & Cowen, L. E. Molecular evolution of antifungal drug resistance. Annu Rev. Microbiol. 71, 753–775 (2017).
pubmed: 28886681 doi: 10.1146/annurev-micro-030117-020345
Lee, Y., Puumala, E., Robbins, N. & Cowen, L. E. Antifungal drug resistance: molecular mechanisms in Candida albicans and beyond. Chem. Rev. 121, 3390–3411 (2021).
pubmed: 32441527 doi: 10.1021/acs.chemrev.0c00199
Uno, N., Li, Z., Avery, L., Sfeir, M. M. & Liu, C. CRISPR gel: a one-pot biosensing platform for rapid and sensitive detection of HIV viral RNA. Anal. Chim. Acta 1262, 341258 (2023).
pubmed: 37179057 pmcid: 10187225 doi: 10.1016/j.aca.2023.341258
Ding, X. et al. Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay. Nat. Commun. 11, 4711 (2020).
pubmed: 32948757 pmcid: 7501862 doi: 10.1038/s41467-020-18575-6
Feng, W. et al. Integrating reverse transcription recombinase polymerase amplification with CRISPR technology for the one-tube assay of RNA. Anal. Chem. 93, 12808–12816 (2021).
pubmed: 34506127 doi: 10.1021/acs.analchem.1c03456
Joung, J. et al. Detection of SARS-CoV-2 with SHERLOCK one-pot testing. N. Engl. J. Med. 383, 1492–1494 (2020).
pubmed: 32937062 doi: 10.1056/NEJMc2026172
Ma, Y. et al. Efficient magnetic enrichment cascade single-step RPA-CRISPR/Cas12a assay for rapid and ultrasensitive detection of Staphylococcus aureus in food samples. J. Hazard Mater. 465, 133494 (2024).
pubmed: 38228008 doi: 10.1016/j.jhazmat.2024.133494
Lin, M. et al. Glycerol additive boosts 100-fold sensitivity enhancement for one-pot RPA-CRISPR/Cas12a assay. Anal. Chem. 94, 8277–8284 (2022).
pubmed: 35635176 doi: 10.1021/acs.analchem.2c00616
Yin, K. et al. Dynamic aqueous multiphase reaction system for one-pot CRISPR-Cas12a-based ultrasensitive and quantitative molecular diagnosis. Anal. Chem. 92, 8561–8568 (2020).
pubmed: 32390420 pmcid: 7588651 doi: 10.1021/acs.analchem.0c01459
Li, J., Macdonald, J. & von Stetten, F. Review: a comprehensive summary of a decade development of the recombinase polymerase amplification. Analyst 144, 31–67 (2018).
pubmed: 30426974 doi: 10.1039/C8AN01621F
Yan, L. et al. Isothermal amplified detection of DNA and RNA. Mol. Biosyst. 10, 970–1003 (2014).
Daher, R. K., Stewart, G., Boissinot, M. & Bergeron, M. G. Recombinase polymerase amplification for diagnostic applications. Clin. Chem. 62, 947–958 (2016).
pubmed: 27160000 pmcid: 7108464 doi: 10.1373/clinchem.2015.245829
James, A.S. & Alawneh, J.I. COVID-19 infection diagnosis: potential impact of isothermal amplification technology to reduce community transmission of SARS-CoV-2. Diagnostics 10, 399 (2020).
Piepenburg, O., Williams, C. H., Stemple, D. L. & Armes, N. A. DNA detection using recombination proteins. PLoS Biol. 4, e204 (2006).
pubmed: 16756388 pmcid: 1475771 doi: 10.1371/journal.pbio.0040204

Auteurs

Shimei Shen (S)

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Department of Dermatology and Cosmetology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, China.
The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.
Department of Clinical Laboratory Medicine, Chongqing Red Cross Hospital (Jiangbei District People's Hospital), Chongqing, China.

Wen Wang (W)

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Department of Laboratory Medicine, The Second Affiliated Hospital of Chengdu Medical College (Nuclear Industry 416 Hospital), Chengdu, China.

Yuanyan Ma (Y)

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

Shilei Wang (S)

Department of Dermatology and Cosmetology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, China.

Shaocheng Zhang (S)

Department of Laboratory Medicine, The Second Affiliated Hospital of Chengdu Medical College (Nuclear Industry 416 Hospital), Chengdu, China.
School of Clinical Medicine, Chengdu Medical College, Chengdu, China.
ORCID: 0000-0003-3620-1616

Xuefei Cai (X)

The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

Liang Chen (L)

Department of Pharmacy Practice, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, Buffalo, NY, USA.
ORCID: 0000-0001-5845-2235

Jin Zhang (J)

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

Yalan Li (Y)

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

Xiaoli Wu (X)

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

Jie Wei (J)

Department of Clinical Laboratory, Zhuhai People's Hospital (Zhuhai Hospital Affiliated with Jinan University), Zhuhai, China.

Yanan Zhao (Y)

Department of Pharmacy Practice, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, Buffalo, NY, USA. yananzh@buffalo.edu.

Ailong Huang (A)

The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China. ahuang@cqmu.edu.cn.
ORCID: 0000-0003-0148-7423

Siqiang Niu (S)

Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China. siqiangniu@cqmu.edu.cn.

Deqiang Wang (D)

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), Department of Laboratory Medicine, Chongqing Medical University, Chongqing, China. wangdq333@cqmu.edu.cn.
The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China. wangdq333@cqmu.edu.cn.
Western (Chongqing) Collaborative Innovation Center for Intelligent Diagnostics and Digital Medicine, Chongqing National Biomedicine Industry Park, Chongqing, China. wangdq333@cqmu.edu.cn.
ORCID: 0009-0004-0643-4376

Articles similaires

[Redispensing of expensive oral anticancer medicines: a practical application].

Lisanne N van Merendonk, Kübra Akgöl, Bastiaan Nuijen
1.00
Humans Antineoplastic Agents Administration, Oral Drug Costs Counterfeit Drugs

Smoking Cessation and Incident Cardiovascular Disease.

Jun Hwan Cho, Seung Yong Shin, Hoseob Kim et al.
1.00
Humans Male Smoking Cessation Cardiovascular Diseases Female

Evaluation of Low-Value Services Across Major Medicare Advantage Insurers and Traditional Medicare.

Ciara Duggan, Adam L Beckman, Ishani Ganguli et al.
1.00
Humans United States Aged Cross-Sectional Studies Medicare Part C

Effectiveness of Virtual Yoga for Chronic Low Back Pain: A Randomized Clinical Trial.

Hallie Tankha, Devyn Gaskins, Amanda Shallcross et al.
1.00
Humans Yoga Low Back Pain Female Male

Classifications MeSH

questionsmedicales.fr › Eucaryotes › Champignons › Levures › Candida › Candida albicans › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Glucides › Polyosides › Chitine › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Systèmes CRISPR-Cas › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ADN › ADN fongique › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Environnement et santé publique › Santé publique › Méthodes épidémiologiques › Statistiques comme sujet › Sensibilité et spécificité › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Techniques d'investigation › Irrigation thérapeutique › Lavage bronchoalvéolaire › Liquide de lavage bronchoalvéolaire › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Hydrolases › Esterases › Endonucleases › Endodeoxyribonucleases › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Infection › Infections bactériennes et mycoses › Mycoses › Candidose › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines bactériennes › Affinity molecular assay for detecting Candida...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines associées aux CRISPR › Affinity molecular assay for detecting Candida...