Identification of Fungal Species and Detection of Azole-Resistance Mutations in the Aspergillus fumigatus cyp51A Gene at a South Korean Hospital
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: With changing fungal epidemiology and azole resistance in Aspergillus species, identifying fungal species and susceptibilitypatterns is crucial to the management of aspergillosis and mucormycosis. The objectives of this study were to evaluate performanceof panfungal polymerase chain reaction (PCR) assays on formalin-fixed paraffin embedded (FFPE) samples in theidentification of fungal species and in the detection of azole-resistance mutations in the Aspergillus fumigatus cyp51A gene at aSouth Korean hospital. Materials and Methods: A total of 75 FFPE specimens with a histopathological diagnosis of aspergillosis or mucormycosis wereidentified during the 10-year study period (2006–2015). After deparaffinization and DNA extraction, panfungal PCR assays wereconducted on FFPE samples for fungal species identification. The identified fungal species were compared with histopathologicaldiagnosis. On samples identified as A. fumigatus, sequencing to identify frequent mutations in the cyp51A gene [tandem repeat46 (TR46), L98H, and M220 alterations] that confer azole resistance was performed. Results: Specific fungal DNA was identified in 31 (41.3%) FFPE samples, and of these, 16 samples of specific fungal DNA were inaccord with a histopathological diagnosis of aspergillosis or mucormycosis; 15 samples had discordant histopathology and PCRresults. No azole-mediating cyp51A gene mutation was noted among nine cases of aspergillosis. Moreover, no cyp51A mutationswere identified among three cases with history of prior azole use. Conclusion: Panfungal PCR assay with FFPE samples may provide additional information of use to fungal species identification. Noazole-resistance mediating mutations in the A. fumigatus cyp51A gene were identified among FFPE samples during study period.
Abstract
Purpose: With changing fungal epidemiology and azole resistance in Aspergillus species, identifying fungal species and susceptibilitypatterns is crucial to the management of aspergillosis and mucormycosis. The objectives of this study were to evaluate performanceof panfungal polymerase chain reaction (PCR) assays on formalin-fixed paraffin embedded (FFPE) samples in theidentification of fungal species and in the detection of azole-resistance mutations in the Aspergillus fumigatus cyp51A gene at aSouth Korean hospital. Materials and Methods: A total of 75 FFPE specimens with a histopathological diagnosis of aspergillosis or mucormycosis wereidentified during the 10-year study period (2006–2015). After deparaffinization and DNA extraction, panfungal PCR assays wereconducted on FFPE samples for fungal species identification. The identified fungal species were compared with histopathologicaldiagnosis. On samples identified as A. fumigatus, sequencing to identify frequent mutations in the cyp51A gene [tandem repeat46 (TR46), L98H, and M220 alterations] that confer azole resistance was performed. Results: Specific fungal DNA was identified in 31 (41.3%) FFPE samples, and of these, 16 samples of specific fungal DNA were inaccord with a histopathological diagnosis of aspergillosis or mucormycosis; 15 samples had discordant histopathology and PCRresults. No azole-mediating cyp51A gene mutation was noted among nine cases of aspergillosis. Moreover, no cyp51A mutationswere identified among three cases with history of prior azole use. Conclusion: Panfungal PCR assay with FFPE samples may provide additional information of use to fungal species identification. Noazole-resistance mediating mutations in the A. fumigatus cyp51A gene were identified among FFPE samples during study period.
참고문헌 (33)
- Buil JB, "Molecular detection of azole-resistant Aspergillus fumigatus in clinical samples", Front Microbiol, 9, pp.515~ (2018)
- Kauffman CA, "Zygomycosis : an emerging fungal infection with new options for management", Curr Infect Dis Rep, 9, pp.435~440 (2007)
- Patterson TF, "Practice guidelines for the diagnosis and management of aspergillosis : 2016 update by the Infectious Diseases Society of America", Clin Infect Dis, 63, pp.e1~e60 (2016)
- Cho SY, "Epidemiology and antifungal susceptibility profile of aspergillus species : comparison between environmental and clinical isolates from patients with hematologic malignancies", J Clin Microbiol, 57, pp.e02023~ (2019)
- Huh K, "Serious fungal infections in Korea", Eur J Clin Microbiol Infect Dis, 36, pp.957~963 (2017)
- 김성한, "침습성 폐 아스페르길루스증의 역학과 임상상: 전국 다기관 연구", Infection and Chemotherapy, 44(4), pp.282~288 (2012), 대한감염학회
- Lestrade PP, "Voriconazole resistance and mortality in invasive aspergillosis : a multicenter retrospective cohort study", Clin Infect Dis, 68, pp.1463~1471 (2019)
- Dudakova A, "Molecular tools for the detection and deduction of Azole Antifungal drug resistance phenotypes in Aspergillus species", Clin Microbiol Rev, 30, pp.1065~1091 (2017)
- Wiederhold NP, "First detection of TR34 L98H and TR46 Y121F T289A Cyp51 mutations in Aspergillus fumigatus Isolates in the United States", J Clin Microbiol, 54, pp.168~171 (2016)
- Chowdhary A, "Azole-resistant aspergillosis : epidemiology, molecular mechanisms, and treatment", J Infect Dis, 216, pp.S436~S444 (2017)
- Salehi E, "Discrimination of aspergillosis, mucormycosis, fusariosis, and scedosporiosis in formalin-fixed paraffin-embedded tissue specimens by use of multiple real-time quantitative PCR assays", J Clin Microbiol, 54, pp.2798~2803 (2016)
- de Groot T, "Genotyping of aspergillus fumigatus in formalin-fixed paraffinembedded tissues and serum samples from patients with invasive aspergillosis", Front Cell Infect Microbiol, 8, pp.377~ (2018)
- McCarthy MW, "Future research priorities in fungal resistance", J Infect Dis, 216, pp.S484~S492 (2017)
- Babouee Flury B, "Performances of two different panfungal PCRs to detect mould DNA in formalin-fixed paraffin-embedded tissue : what are the limiting factors", BMC Infect Dis, 14, pp.692~ (2014)
- Spiess B, "Incidence of Cyp51 A key mutations in Aspergillus fumigatus-a study on primary clinical samples of immunocompromised patients in the period of 1995-2013", PLoS One, 9, pp.e103113~ (2014)
- Spiess B, "Development of novel PCR assays to detect azole resistancemediating mutations of the Aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients", Antimicrob Agents Chemother, 56, pp.3905~3910 (2012)
- Lee HJ, "TR34", L98H mutation in CYP51A gene in Aspergillus fumigatus clinical isolates during posaconazole prophylaxis : first case in Korea, 2018, pp.183, 731~736
- Bialek R, "PCR based identification and discrimination of agents of mucormycosis and aspergillosis in paraffin wax embedded tissue", J Clin Pathol, 58, pp.1180~1184 (2005)
- Hayden RT, "In situ hybridization for the identification of filamentous fungi in tissue section", Diagn Mol Pathol, 11, pp.119~126 (2002)
- Kutlu M, "Pulmonary mucormycosis due to Lichtheimia ramosa in a patient with HIV infection", Mycopathologia, 178, pp.111~115 (2014)
- Mouronte-Roibás C, "Lichtheimia ramosa : a fatal case of mucormycosis", Can Respir J, 2016, pp.2178218~ (2016)
- de Chaumont A, "Mucormycosis : a rare complication of an amputation", Ann Vasc Surg, 28, pp.1035e15~1035e19 (2014)
- Ibrahim AS, "Caspofungin inhibits Rhizopus oryzae 1, 3-beta-D-glucan synthase, lowers burden in brain measured by quantitative PCR, and improves survival at a low but not a high dose during murine disseminated zygomycosis", Antimicrob Agents Chemother, 49, pp.721~727 (2005)
- Makimura K, "Detection of a wide range of medically important fungi by the polymerase chain reaction", J Med Microbiol, 40, pp.358~364 (1994)
- Bialek R, "Detection of Cryptococcus neoformans DNA in tissue samples by nested and real-time PCR assays", Clin Diagn Lab Immunol, 9, pp.461~469 (2002)
- Gonzalez CE, "Disseminated zygomycosis in a neutropenic patient : successful treatment with amphotericin B lipid complex and granulocyte colony-stimulating factor", Clin Infect Dis, 24, pp.192~196 (1997)
- Chamilos G, "Delaying amphotericin B-based frontline therapy significantly increases mortality among patients with hematologic malignancy who have zygomycosis", Clin Infect Dis, 47, pp.503~509 (2008)
- Bueid A, "Azole antifungal resistance in Aspergillus fumigatus : 2008 and 2009", J Antimicrob Chemother, 65, pp.2116~2118 (2010)
- Howard SJ, "Multi-azole resistance in Aspergillus fumigatus", Int J Antimicrob Agents, 28, pp.450~453 (2006)
- Lockhart SR, "Azole resistance in Aspergillus fumigatus isolates from the ARTEMIS global surveillance study is primarily due to the TR", L98H mutation in the cyp51A gene, 2011, pp.55, 4465~4468
- van der Linden JW, "Clinical implications of azole resistance in Aspergillus fumigatus, The Netherlands, 2007-2009", Emerg Infect Dis, 17, pp.1846~1854 (2011)
- Chowdhary A, "Multi-azole-resistant Aspergillus fumigatus in the environment in Tanzania", J Antimicrob Chemother, 69, pp.2979~2983 (2014)
- Chowdhary A, "Clonal expansion and emergence of environmental multipletriazole-resistant Aspergillus fumigatus strains carrying the TR₃₄", L98H mutations in the cyp51A gene in India, 2012, pp.7, e52871~