皮肤癣菌的分子鉴定技术研究进展

摘要/Abstract

摘要： 皮肤癣菌是临床最常见的真菌之一，目前已报道的50余种皮肤癣菌中有20余种对人类有致病性。新兴的分子鉴定技术，具有快速、简便、特异的优点，能够克服传统的直接镜检和病原菌培养耗时费力的局限性，帮助皮肤癣菌病的快速精准诊断和合理用药。该文回顾近二十年来国内外文献，从皮肤癣菌标记基因和分类学进展、基于PCR的分子鉴定技术、恒温核酸扩增技术和MALDI-TOF MS技术这四个方面，介绍皮肤癣菌的分子鉴定技术的研究进展。

关键词: 皮肤癣菌病, PCR, 恒温核酸扩增技术, MALDI-TOF MS

中图分类号:

R379.2

石媛漾, 潘炜华, 廖万清, 陈显振, 蔡刘阳. 皮肤癣菌的分子鉴定技术研究进展[J]. 中国真菌学杂志, 2022, 17(4): 330-334.

参考文献

[1] HAVLICKOVA B, CZAIKA V A, FRIEDRICH M.Epidemiological trends in skin mycoses worldwide[J].Mycoses, 2008, 51(Suppl 4):2-15.
[2] 刘维达, 梁官钊, 张美洁, 等.医学真菌分类学最新进展[J].中国真菌学杂志, 2021, 16(1):1-5.
[3] KIZNY GORDON A, MCIVER C, KIM M, et al.Clinical application of a molecular assay for the detection of dermatophytosis and a novel non-invasive sampling technique[J].Pathology, 2016, 48(7):720-726.
[4] JENSEN R H, ARENDRUP M C.Molecular diagnosis of dermatophyte infections[J].Curr Opin Infect Dis, 2012, 25(2):126-134.
[5] VERRIER J, MONOD M.Diagnosis of dermatophytosis using molecular biology[J].Mycopathologia, 2017, 182(1-2):193-202.
[6] SPILIOPOULOU A, BARTZAVALI C, JELASTOPULU E, et al.Evaluation of a commercial PCR test for the diagnosis of dermatophyte nail infections[J].J Med Microbiol, 2015, 64(Pt 1):25-31.
[7] CHOLLET A, CATTIN V, FRATTI M, et al.Which fungus originally was Trichophyton mentagrophytes?Historical review and illustration by a clinical case[J].Mycopathologia, 2015, 180(1-2):1-5.
[8] GRäSER Y, DE HOOG S, SUMMERBELL R C.Dermatophytes:recognizing species of clonal fungi[J].Med Mycol, 2006, 44(3):199-209.
[9] HIRAI A, KANO R, NAKAMURA Y, et al.Molecular taxonomy of dermatophytes and related fungi by chitin synthase 1(CHS1) gene sequences[J].Antonie van Leeuwenhoek, 2003, 83(1):11-20.
[10] KANO R, HIRAI A, MURAMATSU M, et al.Direct detection of dermatophytes in skin samples based on sequences of the chitin synthase 1(CHS1) gene[J].J Vet Med Sci, 2003, 65(2):267-270.
[11] VERRIER J, KRäHENBüHL L, BONTEMS O, et al.Dermatophyte identification in skin and hair samples using a simple and reliable nested polymerase chain reaction assay[J].Br J Dermatol, 2013, 168(2):295-301.
[12] BEIFUSS B, BEZOLD G, GOTTLöBER P, et al.Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer[J].Mycoses, 2011, 54(2):137-145.
[13] BRASCH J, BECK-JENDROSCHEK V, GLäSER R.Fast and sensitive detection of Trichophyton rubrum in superficial tinea and onychomycosis by use of a direct polymerase chain reaction assay[J].Mycoses, 2011, 54(5):e313-7.
[14] REZAEI-MATEHKOLAEI A, MAKIMURA K, DE HOOG G S, et al.Discrimination of Trichophyton tonsurans and Trichophyton equinum by PCR-RFLP and by β-tubulin and translation elongation factor 1-α sequencing[J].Med Mycol, 2012, 50(7):760-764.
[15] REZAEI-MATEHKOLAEI A, MIRHENDI H, MAKIMURA K, et al.Nucleotide sequence analysis of beta tubulin gene in a wide range of dermatophytes[J].Med Mycol, 2014, 52(7):674-688.
[16] MIRHENDI H, MAKIMURA K, DE HOOG G S, et al.Translation elongation factor 1-α gene as a potential taxonomic and identification marker in dermatophytes[J].Med Mycol, 2015, 53(3):215-224.
[17] AHMADI B, MIRHENDI H, MAKIMURA K, et al.Phylogenetic analysis of dermatophyte species using DNA sequence polymorphism in calmodulin gene[J].Med Mycol, 2016, 54(5):500-514.
[18] REZAEI-MATEHKOLAEI A, MAKIMURA K, DE HOOG G S, et al.Multilocus differentiation of the related dermatophytes Microsporum canis, Microsporum ferrugineum and Microsporum audouinii[J].J Med Microbiol, 2012, 61(Pt 1):57-63.
[19] HAYETTE M P, SACHELI R.Dermatophytosis, trends in epidemiology and diagnostic approach[J].Curr Fungal Infect Rep, 2015, 9(3):1-16.
[20] BERGMANS A M, VAN DER ENT M, KLAASSEN A, et al.Evaluation of a single-tube real-time PCR for detection and identification of 11 dermatophyte species in clinical material[J].Clin Microbiol Infect, 2010, 16(6):704-710.
[21] ALEXANDER C L, SHANKLAND G S, CARMAN W, et al.Introduction of a dermatophyte polymerase chain reaction assay to the diagnostic mycology service in Scotland[J].Br J Dermatol, 2011, 164(5):966-972.
[22] WISSELINK G J, VAN ZANTEN E, KOOISTRA-SMID A M.Trapped in keratin;a comparison of dermatophyte detection in nail, skin and hair samples directly from clinical samples using culture and real-time PCR[J].J Microbiol Methods, 2011, 85(1):62-66.
[23] MIYAJIMA Y, SATOH K, UCHIDA T, et al.Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes[J].J Dermatol Sci, 2013, 69(3):229-235.
[24] VERRIER J, BONTEMS O, BAUDRAZ-ROSSELET F, et al.Oral terbinafine and itraconazole treatments against dermatophytes appear not to favor the establishment of Fusarium spp.in nail[J].Dermatology, 2014, 228(3):225-232.
[25] GARG J, TILAK R, SINGH S, et al.Evaluation of pan-dermatophyte nested PCR in diagnosis of onychomycosis[J].J Clin Microbiol, 2007, 45(10):3443-3445.
[26] GARG J, TILAK R, GARG A, et al.Rapid detection of dermatophytes from skin and hair[J].BMC Res Notes, 2009, 2:60.DOI:10.1186/1756-0500-2-60.
[27] CAFARCHIA C, GASSER R B, FIGUEREDO L A, et al.An improved molecular diagnostic assay for canine and feline dermatophytosis[J].Med Mycol, 2013, 51(2):136-143.
[28] PETINATAUD D, BERGER S, CONTET-AUDONNEAU N, et al.Molecular diagnosis of onychomycosis[J].J Mycol Med, 2014, 24(4):287-295.
[29] ARABATZIS M, BRUIJNESTEIJN VAN COPPENRAET L E, KUIJPER E J, et al.Diagnosis of common dermatophyte infections by a novel multiplex real-time polymerase chain reaction detection/identification scheme[J].Br J Dermatol, 2007, 157(4):681-689.
[30] BRILLOWSKA-DABROWSKA A, SAUNTE D M, ARENDRUP M C.Five-hour diagnosis of dermatophyte nail infections with specific detection of Trichophyton rubrum[J].J Clin Microbiol, 2007, 45(4):1200-1204.
[31] ELAVARASHI E, KINDO A J, KALYANI J.Optimization of PCR-RFLP directly from the skin and nails in cases of dermatophytosis, targeting the ITS and the 18S ribosomal DNA regions[J].JCDR, 2013, 7(4):646-651.
[32] VERRIER J, PRONINA M, PETER C, et al.Identification of infectious agents in onychomycoses by PCR-terminal restriction fragment length polymorphism[J].J Clin Microbiol, 2012, 50(3):553-561.
[33] RASMUSSEN H B.Restriction fragment length polymorphism analysis of PCR-amplified fragments (PCR-RFLP) and gel electrophoresis-valuable tool for genotyping and genetic fingerprinting[M/OL]//MAGDELDIN S.Gel electrophoresis-principles and basics.London:IntechOpen, 2012.https://doi.org/10.5772/37724.
[34] BODULEV O L, SAKHAROV I Y.Isothermal nucleic acid amplification techniques and their use in bioanalysis[J].Biochemistry (Mosc), 2020, 85(2):147-166.
[35] NURUL NAJIAN A B, FOO P C, ISMAIL N, et al.Probe-specific loop-mediated isothermal amplification magnetogenosensor assay for rapid and specific detection of pathogenic Leptospira[J].Mol Cell Probes, 2019, 44:63-68.DOI:10.1016/j.mcp.2019.03.001.
[36] SHCHIT I Y, IGNATOV K B, KUDRYAVTSEVA T Y, et al.The use of loop-mediated isothermal DNA amplification for the detection and identification of the anthrax pathogen[J].Mol Gen Microbiol Virol, 2017, 32(2):100-108.
[37] PöSCHL B, WANEESORN J, THEKISOE O, et al.Comparative diagnosis of malaria infections by microscopy, nested PCR, and LAMP in northern Thailand[J].Am J Trop Med Hyg, 2010, 83(1):56-60.
[38] MA Y, DAI X, HONG T, et al.A NASBA on microgel-tethered molecular-beacon microarray for real-time microbial molecular diagnostics[J].Analyst, 2016, 142(1):147-155.
[39] REID M S, LE X C, ZHANG H.Exponential isothermal amplification of nucleic acids and assays for proteins, cells, small molecules, and enzyme activities:an EXPAR example[J].Angew Chem Int Ed Engl, 2018, 57(37):11856-11866.
[40] LIU H, ZHANG L, XU Y, et al.Sandwich immunoassay coupled with isothermal exponential amplification reaction:An ultrasensitive approach for determination of tumor marker MUC1[J].Talanta, 2019, 204:248-254.DOI:10.1016/j.talanta.2019.06.001.
[41] LOBATO I M, O'SULLIVAN C K.Recombinase polymerase amplification:basics, applications and recent advances[J].Trends Analyt Chem, 2018, 98:19-35.DOI:10.1016/j.trac.2017.10.015.
[42] GOO N I, KIM D E.Rolling circle amplification as isothermal gene amplification in molecular diagnostics[J].Biochip J, 2016, 10(4):262-271.
[43] DENG S, ZHOU Z, DE HOOG G S, et al.Evaluation of two molecular techniques for rapid detection of the main dermatophytic agents of tinea capitis[J].Br J Dermatol, 2015, 173(6):1494-500.
[44] DIRKS R M, PIERCE N A.Triggered amplification by hybridization chain reaction[J].Proc Natl Acad Sci U S A, 2004, 101(43):15275-15278.
[45] MIAO P, TANG Y, YIN J.MicroRNA detection based on analyte triggered nanoparticle localization on a tetrahedral DNA modified electrode followed by hybridization chain reaction dual amplification[J].Chem Commun (Camb), 2015, 51(86):15629-15632.
[46] YAN L, ZHOU J, ZHENG Y, et al.Isothermal amplified detection of DNA and RNA[J].Mol Biosyst, 2014, 10(5):970-1003.
[47] KIESLING T, COX K, DAVIDSON E A, et al.Sequence specific detection of DNA using nicking endonuclease signal amplification (NESA)[J].Nucleic Acids Res, 2007, 35(18):e117.
[48] BERTOLI G, CAVA C, CASTIGLIONI I.MicroRNAs:new biomarkers for diagnosis, prognosis, therapy prediction and therapeutic tools for breast cancer[J].Theranostics, 2015, 5(10):1122-1143.
[49] NENOFF P, ERHARD M, SIMON J C, et al.MALDI-TOF mass spectrometry-a rapid method for the identification of dermatophyte species[J].Med Mycol, 2013, 51(1):17-24.
[50] TORRES-SANGIAO E, LEAL RODRIGUEZ C, GARCíA-RIESTRA C.Application and perspectives of MALDI-TOF mass spectrometry in clinical microbiology laboratories[J].Microorganisms, 2021, 9(7):1539.
[51] L'OLLIVIER C, RANQUE S.MALDI-TOF-based dermatophyte identification[J].Mycopathologia, 2017, 182(1-2):183-192.
[52] HOU T Y, CHIANG-NI C, TENG S H.Current status of MALDI-TOF mass spectrometry in clinical microbiology[J].J Food Drug Anal, 2019, 27(2):404-414.