A novel rRNA hybridization-based approach to rapid, accurate Candida identification directly from blood culture.
A novel rRNA hybridization-based approach to rapid, accurate Candida identification directly from blood culture.
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一种基于 rRNA 杂交的新型方法，可直接从血培养物中快速、准确地鉴定念珠菌。
DOI：
10.1093/mmy/myac065
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发表时间：
2022
期刊：
影响因子：
2.9
通讯作者：
Bhattacharyya,RobyP
中科院分区：
文献类型：
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作者：
Matzko,MichelleE;Sephton-Clark,PoppyCS;Young,EleanorL;Jhaveri,TulipA;Martinsen,MelanieA;Mojica,Evan;Boykin,Rich;Pierce,VirginiaM;Cuomo,ChristinaA;Bhattacharyya,RobyP
侵袭性真菌感染越来越普遍，具有高发病率和死亡率，但真菌诊断落后于细菌诊断在快速确定病原。我们之前设计了一种基于荧光杂交的检测方法，可以在几小时内直接从血培养瓶中鉴定细菌，称为基于系统发育的rrna菌株鉴定（phfirst - id）。在这里，我们采用这种方法明确地鉴定了11种常见的致病性念珠菌，包括c。Auris，实验室培养的准确度为100%（33株菌株中的33株在参考小组中，加上33株在验证小组中测试的额外分离株中的33株）。在对来自两家医院的62个连续阳性临床血液培养物的初步研究中，革兰氏染色上显示酵母菌，candiaphirst - id与11个物种参考小组中59个样本中的58个样本的临床实验室结果相匹配，没有对3个样本外的物种进行错误分类。它还在这62个标本中的2个标本中检测到混合念珠菌，包括一个分类不一致的标本，这些标本未被标准临床微生物学工作流程识别；在每种情况下，两种物种的存在都得到了临床和实验数据的验证。最后，在三个同时生长细菌和酵母的样本中，我们将先前的细菌探针集与新的假丝酵母探针集配对，使用phfirst - id检测两种病原体类型。这种简单、可靠的检测方法可以在数小时内直接从血液培养瓶中提供准确的细菌鉴定，并且概念方法有望在单一工作流程中进行泛微生物鉴定。念珠菌感染引起相当大的发病率和死亡率，但缓慢的诊断延误了识别，恶化了患者的预后。我们开发并验证了一种新的分子方法，可以比标准工作流程更快地从血培养中准确识别念珠菌。
Invasive fungal infections are increasingly common and carry high morbidity and mortality, yet fungal diagnostics lag behind bacterial diagnostics in rapidly identifying the causal pathogen. We previously devised a fluorescent hybridization-based assay to identify bacteria within hours directly from blood culture bottles without subculture, called phylogeny-informed rRNA-based strain identification (Phirst-ID). Here, we adapt this approach to unambiguously identify 11 common pathogenicCandidaspecies, includingC. auris, with 100% accuracy from laboratory culture (33 of 33 strains in a reference panel, plus 33 of 33 additional isolates tested in a validation panel). In a pilot study on 62 consecutive positive clinical blood cultures from two hospitals that showed yeast on Gram stain,CandidaPhirst-ID matched the clinical laboratory result for 58 of 59 specimens represented in the 11-species reference panel, without misclassifying the 3 off-panel species. It also detected mixedCandidaspecies in 2 of these 62 specimens, including the one discordant classification, that were not identified by standard clinical microbiology workflows; in each case the presence of both species was validated by both clinical and experimental data. Finally, in three specimens that grew both bacteria and yeast, we paired our prior bacterial probeset with this newCandidaprobeset to detect both pathogen types using Phirst-ID. This simple, robust assay can provide accurateCandidaidentification within hours directly from blood culture bottles, and the conceptual approach holds promise for pan-microbial identification in a single workflow.Lay SummaryCandidabloodstream infections cause considerable morbidity and mortality, yet slow diagnostics delay recognition, worsening patient outcomes. We develop and validate a novel molecular approach to accurately identifyCandidaspecies directly from blood culture one day faster than standard workflows.