阿根廷龈下样本中白色念珠菌和都柏林念珠菌gDNA的提取。不同方法的评估。

Verónica A Dubois, Pablo A Salgado, Laura A Gliosca, Susana L Molgatini
{"title":"阿根廷龈下样本中白色念珠菌和都柏林念珠菌gDNA的提取。不同方法的评估。","authors":"Verónica A Dubois,&nbsp;Pablo A Salgado,&nbsp;Laura A Gliosca,&nbsp;Susana L Molgatini","doi":"10.54589/aol.36/2/78","DOIUrl":null,"url":null,"abstract":"<p><p>The oral cavity constitutes a unique ecosystem with highly variable ecological niches that harbor a great variety of microorganisms, including yeasts. Molecular methods are currently considered the gold standard for identifying species, although they involve limitations associated with the disruption of yeast cell walls to release the genomic DNA (gDNA) for amplification.</p><p><strong>Aim: </strong>The aim of this study was to compare the performance of different methods for extracting gDNA from Candida albicans and Candida dubliniensis, subsequently amplifying DNA by PCR.</p><p><strong>Materials and method: </strong>Fifty-two isolates (16 C. albicans and 36 C. dubliniensis) were obtained from subgingival biofilm of HIV+ patients with clinical signs of periodontal disease. The study evaluated 6 gDNA extraction methods and two PCR amplification methods. Furthermore, the presence of alleles of HWP1 gene was determined in C. albicans.</p><p><strong>Results: </strong>Comparisons of six methods show statistically significant differences (p <0.001) except for C. albicans in two of them. For C. dubliniensis, statistical differences were observed in all comparisons. Commercial methods were more efficient for concentrating gDNA than in-house methods, and both PCRs were effective. Ten heterozygous C. albicans isolates for this allele were positive for the HWP1-1 / HWP1-2 allele, one was homozygous for Wild Type HWP1-1 allele, and 5 were homozygous for novel/rare HWP1-2 allele.</p><p><strong>Conclusions: </strong>This study aims to provide simple, inexpensive strategies for phenotypic identification and molecular confirmation of Candida albicans and Candida dubliniensis for non-reference laboratories with low complexity and/or low budgets.</p>","PeriodicalId":93853,"journal":{"name":"Acta odontologica latinoamericana : AOL","volume":"36 2","pages":"78-85"},"PeriodicalIF":0.0000,"publicationDate":"2023-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e5/82/1852-4834-36-2-78.PMC10557081.pdf","citationCount":"0","resultStr":"{\"title\":\"gDNA extraction from Candida albicans and Candida dubliniensis in subgingival samples in Argentina. Evaluation of different methods.\",\"authors\":\"Verónica A Dubois,&nbsp;Pablo A Salgado,&nbsp;Laura A Gliosca,&nbsp;Susana L Molgatini\",\"doi\":\"10.54589/aol.36/2/78\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The oral cavity constitutes a unique ecosystem with highly variable ecological niches that harbor a great variety of microorganisms, including yeasts. Molecular methods are currently considered the gold standard for identifying species, although they involve limitations associated with the disruption of yeast cell walls to release the genomic DNA (gDNA) for amplification.</p><p><strong>Aim: </strong>The aim of this study was to compare the performance of different methods for extracting gDNA from Candida albicans and Candida dubliniensis, subsequently amplifying DNA by PCR.</p><p><strong>Materials and method: </strong>Fifty-two isolates (16 C. albicans and 36 C. dubliniensis) were obtained from subgingival biofilm of HIV+ patients with clinical signs of periodontal disease. The study evaluated 6 gDNA extraction methods and two PCR amplification methods. Furthermore, the presence of alleles of HWP1 gene was determined in C. albicans.</p><p><strong>Results: </strong>Comparisons of six methods show statistically significant differences (p <0.001) except for C. albicans in two of them. For C. dubliniensis, statistical differences were observed in all comparisons. Commercial methods were more efficient for concentrating gDNA than in-house methods, and both PCRs were effective. Ten heterozygous C. albicans isolates for this allele were positive for the HWP1-1 / HWP1-2 allele, one was homozygous for Wild Type HWP1-1 allele, and 5 were homozygous for novel/rare HWP1-2 allele.</p><p><strong>Conclusions: </strong>This study aims to provide simple, inexpensive strategies for phenotypic identification and molecular confirmation of Candida albicans and Candida dubliniensis for non-reference laboratories with low complexity and/or low budgets.</p>\",\"PeriodicalId\":93853,\"journal\":{\"name\":\"Acta odontologica latinoamericana : AOL\",\"volume\":\"36 2\",\"pages\":\"78-85\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-08-31\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e5/82/1852-4834-36-2-78.PMC10557081.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Acta odontologica latinoamericana : AOL\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.54589/aol.36/2/78\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Acta odontologica latinoamericana : AOL","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.54589/aol.36/2/78","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

口腔构成了一个独特的生态系统,具有高度可变的生态位,其中含有多种微生物,包括酵母。分子方法目前被认为是鉴定物种的金标准,尽管它们涉及与破坏酵母细胞壁以释放基因组DNA(gDNA)进行扩增相关的限制。目的:比较不同方法从白色念珠菌和杜氏念珠菌中提取gDNA,然后用PCR扩增DNA的性能。材料和方法:从有牙周病临床症状的HIV+患者的龈下生物膜中分离出52个分离株(16个白色念珠菌和36个杜氏念珠菌)。本研究评估了6种gDNA提取方法和两种PCR扩增方法。此外,在白色念珠菌中检测到HWP1基因等位基因的存在。结果:六种方法的比较显示出统计学上的显著差异(p结论:本研究旨在为低复杂性和/或低预算的非参考实验室提供简单、廉价的白色念珠菌和都柏林念珠菌表型鉴定和分子确认策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

摘要图片

摘要图片

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
gDNA extraction from Candida albicans and Candida dubliniensis in subgingival samples in Argentina. Evaluation of different methods.

The oral cavity constitutes a unique ecosystem with highly variable ecological niches that harbor a great variety of microorganisms, including yeasts. Molecular methods are currently considered the gold standard for identifying species, although they involve limitations associated with the disruption of yeast cell walls to release the genomic DNA (gDNA) for amplification.

Aim: The aim of this study was to compare the performance of different methods for extracting gDNA from Candida albicans and Candida dubliniensis, subsequently amplifying DNA by PCR.

Materials and method: Fifty-two isolates (16 C. albicans and 36 C. dubliniensis) were obtained from subgingival biofilm of HIV+ patients with clinical signs of periodontal disease. The study evaluated 6 gDNA extraction methods and two PCR amplification methods. Furthermore, the presence of alleles of HWP1 gene was determined in C. albicans.

Results: Comparisons of six methods show statistically significant differences (p <0.001) except for C. albicans in two of them. For C. dubliniensis, statistical differences were observed in all comparisons. Commercial methods were more efficient for concentrating gDNA than in-house methods, and both PCRs were effective. Ten heterozygous C. albicans isolates for this allele were positive for the HWP1-1 / HWP1-2 allele, one was homozygous for Wild Type HWP1-1 allele, and 5 were homozygous for novel/rare HWP1-2 allele.

Conclusions: This study aims to provide simple, inexpensive strategies for phenotypic identification and molecular confirmation of Candida albicans and Candida dubliniensis for non-reference laboratories with low complexity and/or low budgets.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
CiteScore
1.00
自引率
0.00%
发文量
0
期刊最新文献
Camouflaged prejudice and the affirmation of skin colour differences: assessment of racism among Brazilian undergraduate dental students. Changes in masseter muscle structure, membrane lipid peroxidation and Ca-ATPase activity as effects of different local anesthetics. Dental caries lesions and impact on quality of life in adolescents living in urban and rural areas. A case study. Effect of two oxygen-inhibiting agents on the surface microhardness of giomer restorative materials. Local application of melatonin associated or not to xenogeneic material, in critical defects of rat calvaria.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1