Study on ERG gene family mutation in fluconazole-resistant vaginal Candida albicans and antimicrobial analysis of Azole drugs

doi:10.3877/cma.j.issn.2095-5820.2022.03.003

Abstract

Abstract:

Objective

To study ERG11, ERG3 and ERG5 gene mutations of Fluconazol-resistant vaginal Candida albicans and the antimicrobial analysis of the isolates against ketoconazole, clotrimazole and miconazole nitrate.

Methods

221 strains of Candida albicans were collected and identified by mass spectrometer. Fluconazole, ketoconazole, clotrimazole and miconazole nitrate were tested by microbroth dilution method, and the minimum inhibitory concentration (MIC) of the four drugs against Candida albicans was obtained. Fluconazole non-sensitive Candida albicans was screened according to CLSI-M27. ERG11, ERG3 and ERG5 genes were amplified after DNA extraction, and bidirectional sequencing was performed, which was compared with the standard sequence (SC5314) and AF069752 in GenBank.

Results

The resistance rate of Candida albicans to fluconazole was 9.05%. Compared with the MIC values of Candida albicans, the MIC values of ketoconazole were lower than those of clotrimazole and miconazole nitrate, with statistical significance (P＜0.05). There was no significant difference between clotrimazole and miconazole nitrate (P＞0.05). ERG11, ERG3 and ERG5 were showed 18, 17 and 12 missense mutations, respectively.

Conclusion

Ketoconazole has stronger antibacterial effect on Candida albicans than metrimazole and miconazole nitrate.. Several missense mutations occurred in ERG11, ERG3 and ERG5 genes of Fluconazol-resistant Candida albicans, and amino acid variation caused by mutations at some sites may be related to drug resistance.

Key words: Candida albicans, ERG genes, Fluconazole, Drug resistance

Qianjun Li, Shuhong Huang, Yonglin Feng, Weihong Lin. Study on ERG gene family mutation in fluconazole-resistant vaginal Candida albicans and antimicrobial analysis of Azole drugs[J]. Chinese Journal of Clinical Laboratory Management(Electronic Edition), 2022, 10(03): 137-141,152.

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Figures/Tables 9

References 13

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引物名称	引物序列（5'-3'）	扩增长度（bp）
ERG11-F	AAGATCATAACTCAATATGGCTA^［6］	1587
ERG11-R	AACACTGAATCGAAAGAAAG	1587
ERG3-F	ATGGATATCGTACTAGAAATTTGTGA^［7］	1161
ERG3-R	TCATTGTTCAACATATTCTCTATCG	1161
ERG5-F	ATGAATTCAACAGAGGTCGATAA^［8］	1554
ERG5-R	CTATAAACTCTTTAATGGGTCTCT	1554

引物名称	引物序列（5'-3'）	扩增长度（bp）
ERG11-F	AAGATCATAACTCAATATGGCTA^［6］	1587
ERG11-R	AACACTGAATCGAAAGAAAG	1587
ERG3-F	ATGGATATCGTACTAGAAATTTGTGA^［7］	1161
ERG3-R	TCATTGTTCAACATATTCTCTATCG	1161
ERG5-F	ATGAATTCAACAGAGGTCGATAA^［8］	1554
ERG5-R	CTATAAACTCTTTAATGGGTCTCT	1554

药物	MIC₅₀	MIC几何均数
酮康唑	3.597	1.817
克霉唑	4.857^a	2.269^a
硝酸咪康唑	5.079^a	2.441^a

药物	MIC₅₀	MIC几何均数
酮康唑	3.597	1.817
克霉唑	4.857^a	2.269^a
硝酸咪康唑	5.079^a	2.441^a

基因	突变菌株序号	变异氨基酸位点	发生突变几率（%）
ERG11	3	A114S	5
	680，688，810	D116E	15
	753	E266D	5
	18	G450E	5
	680，810	G465S	10
	680，810	K128T	10
	2，10	M527I	10
	1，2，10，20，32，34，675，687，703，704，705，714	T123I	60
	1，2，10，18，20，32，34，675，680，687，703，704，705，714，810	Y132H	75
	3	Y257H	5
	3，32，680	终止密码突变	15
ERG3	687	E375K	5
	3，703，741，803	F13L	20
	2	G372E	5
	18，680	G376S	10
	675	L12F	5
	2，3，10，18，34，675，680，687，703，704，705，714，741，753，803，810	V351A	80
	3，703，704	V374G	15
	18，680	V374S	10
	687	Y10N	5
ERG5	1	D503G	5
	1	I505E	5
	2，3，704，810	I505N	20
	10	L504W	5
	3	Q13H	5
	1	Q14P	5
	3	无义1	5
	18，687	无义2	10

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