多重实时荧光定量PCR检测肠出血性大肠杆菌O157:H7

目的 利用多重荧光定量PCR技术,建立一种快速、准确、特异检测肠出血性大肠杆菌O157:H7的定量方法.方法 选取肠出血性大肠杆菌O157:H7编码脂多糖基因(rfbE)和编码鞭毛抗原基因(fliC)作为检测的靶基因,设计引物和TaqMan-MGB探针,探针的5'端分别用FAM和HEX进行荧光标记,3'端标记MGB.优化PCR扩增体系,对多重实时荧光定量PCR方法的特异性、灵敏度、重复性评价,同时进行一定数量临床样本鉴定,与常规方法进行比较.结果 本研究所建立的多重实时荧光定量PCR方法可准确、特异地检测和鉴定肠出血性大肠杆菌O157:H7,能够有效甄别肠出血性大肠杆菌O157:H7与非H7菌株,其他菌株均无阳性结果;该方法的灵敏度可达到10 CFU/ml;定量检测的批间和批内变异系数均小于5%;对66例临床样本进行评价,结果显示15例肠出血性大肠杆菌O157:H7阳性,2例为肠出血性大肠杆菌O157:非H7阳性,其中16例与常规培养法结果符合,符合率达到98.49%.结论 本研究建立的检测肠出血性大肠杆菌O157:H7多重实时荧光定量PCR方法快速,结果准确、可靠,操作简便,为肠出血性大肠杆菌O157:H7的临床诊断、现场流行病学调查和食品安全监测提供了新的鉴定方法.     

实时荧光RT-PCR检测活性单核细胞增生李斯特菌方法的建立

目的 采用逆转录结合实时荧光定量PCR技术,建立一种快速、准确、特异甄别单核细胞增生李斯特菌(Listeria monocytogenes,简称单增李氏菌)死活状态的定量方法.方法 根据单增李氏菌hlyA基因序列设计引物和探针;对实时荧光PCR反应体系进行优化后,提取菌体mRNA,通过随机引物进行逆转录反应;产生的cDNA通过实时荧光定量PCR进行鉴定.进一步评价逆转录结合实时荧光定量PCR方法的特异性、灵敏度、重复性后,对20份模拟双盲样本进行检测.结果 本实验所建立的逆转录结合实时荧光定量PCR方法可准确、特异地检测单增李氏菌,其他菌株和失活的单增李氏菌均无阳性结果出现;该方法检测纯菌和模拟样本的灵敏度分别可达到10 CFU/ml和1000CFU/ml;定量检测的批间和批内的变异系数均小于5%;对20份模拟样本进行检测,其中10份含有活性单增李氏菌样本的检测结果均为阳性,其他含有失活单增李氏菌的5份样本和其他致病菌的5份样本检测结果为阴性.结论 本文建立的检测活性单增李氏菌实时荧光定量PCR方法快速、准确,结果可靠,实用性强,可进行定量分析,为食品安全监测和现场流行病学调查提供较好的分析手段和完整的数据.     

诺如病毒遗传组I型TaqMan—MGB探针实时荧光RT-PCR的研究

目的建立诺如病毒遗传组I型TaqMan-MGB探针实时荧光RT-PCR快速、特异、灵敏的检测方法，为疾病预防控制提供可靠的依据。方法根据GenBank诺如病毒遗传组I型代表株保守序列设计特异引物对和TaqMan-MGB探针，建立一步法实时荧光RT-PCR快速检测反应体系，优化反应条件，评价反应体系的灵敏度、特异性、重复性．并与常规RT—PCR比较。结果诺如病毒遗传组I型TaqMan-MGB探针实时荧光RT-PCR检测时限短。仅1h就出结果。与轮状病毒、腺病毒、星状病毒、甲肝病毒、诺如病毒遗传组Ⅱ型无交叉反应，最低检出下限为100拷贝／反应，比常规RT—PCR灵敏100倍，5份浓度不同的诺如病毒遗传组I型标本重复检测5次。平均Ct值变异系数范围为0．39％-1．02％。结论诺如病毒遗传组I型TaqMan-MGB探针实时荧光RT—PCR快速、特异、灵敏、重复性好，可应用于突发公共卫生应急检测和诺如病毒遗传组I型监测，提高快速检测能力。     

马尔堡、埃博拉病毒双重荧光定量PCR检测方法的建立

目的 建立一种快速、敏感、特异的双重实时荧光定量PCR方法,可同时检测马尔堡病毒和埃博拉病毒.方法 通过序列比对挑选出两种病毒基因组中高度保守的序列,分别设计引物及Taqman探针,两条探针分别标记FAM和Texas Red荧光报告基因,建立双重实时荧光定量PCR反应体系.结果 双重荧光定量PCR方法检测两种病毒阳性标准品的灵敏度分别为30.5拷贝/μl和28.6拷贝/μl,通过检测日本脑炎病毒、黄热病毒、登革热病毒无交叉反应,有较好的灵敏度和特异性.结论 建立了马尔堡、埃博拉病毒双重荧光定量PCR检测方法,实现了两种病毒同时实时定量检测,在传染病防控领域有较好的应用前景.     

肝螺杆菌TaqMan MGB探针实时荧光定量PCR快速检测方法的建立及应用研究

目的 建立特异、敏感、快速检测肝螺杆菌的TaqMan MGB探针实时荧光定量PCR方法.方法 针对肝螺杆菌flaB 基因的保守区设计特异性引物和探针,建立肝螺杆菌TaqMan MGB探针实时荧光定最PCR方检测方法,验证方法的特异性、敏感性和稳定性.对2008-2011年期间采集的1081份临床样本中的肝螺杆菌进行检测,同时进行分离培养和常规PCR检测.结果 建立的TaqMan MGB探针实时荧光定量PCR方法对肝螺杆菌的检测具有高度的特异性,对幽门螺杆菌、空肠弯曲菌、泰泽氏菌、侵肺巴斯德氏菌、大肠埃希菌、铜绿假单胞菌均无交叉反应,检测的灵敏度达8.3拷贝.标准曲线显示各浓度范围内具有良好的线性关系,相关系数为0.999,斜率为-3.227,TaqManMGB探针实时荧光定量PCR效率为100％.对1081份临床样本进行检测,TaqMan MGB探针实时荧光定量PCR和常规PCR均能检出86份肝螺杆菌阳性样本,而细菌分离培养则仅检出4份阳性.结果显示,建立的TaqMan MGB探针实时荧光定量PCR方法比细菌分离培养方法更敏感,能够直接从临床样本中检出肝螺杆菌DNA,检测时间仅为2h.结论 研究建立的TaqMan MGB探针实时荧光定量PCR方法具有可靠、特异、敏感的特点,适用于肝螺杆菌的快速检测.     

荧光定量PCR检测查菲埃立克体

依据查菲埃立克体16SrRNA基因序列设计特异性引物和TaqMan-MGB探针,以克隆的查菲埃立克体16SrRNA基因片段作DNA模板,建立实时荧光定量PCR检测方法。与套式PCR相比较,荧光定量PCR检测的灵敏度是其30倍;用荧光定量PCR检测其他相关立克次体和细菌DNA样本,检出结果为0;对荧光定量PCR检测重复性进行分析,变异系数(CV)批内和批间误差在0·2%~2·0%之间。结果证明本研究建立的荧光定量PCR方法具有种特异性和良好的重复性,可用于检测感染样本中的微量查菲埃立克体DNA。     

应用单一和双重荧光定量PCR法快速检测军团菌

目的 建立单一和双重荧光定量PCR方法分别和同时进行军团菌属及嗜肺军团菌的检测.方法 利用军团菌属16 S rRNA基因和嗜肺军团菌mip基因设计引物和探针,两条基因探针分别标记FAM和HEX,并将相关反应体系和条件进行优化.分别应用单一基因探针(单一荧光定量PCR)和双重基因探针(双重荧光定量PCR)对嗜肺军团菌、非嗜肺军团菌及非军团菌进行检测,并验证两种方法的特异度、敏感度.应用双重荧光定量PCR检测空调水样滤膜样品和DNA提取样品,比较两者结果的一致性.结果 针对军团菌属及嗜肺军团菌,应用荧光定量PCR,16 S rRNA基因和mip基因均能较好的检出,16S rRNA和mip的最低检出限分别为8和10个拷贝.经优化得到了最佳反应体系.单一荧光定量PCR方法所检的8株嗜肺军用菌及4株非嗜肺军团菌16 S rRNA基因均为阳性,嗜肺军团菌mip基因阳性,非嗜肺军团菌mip基因阴性.双重荧光定量PCR方法所检的23株嗜肺军团菌中有2株为假阴性,9株非嗜肺军团菌和非军团菌属中有1株为假阳性.49份空调水样滤膜直接检测和提取DNA后检测的结果一致,其中26份水样军团菌阳性,20份为嗜肺军团菌,6份为非嗜肺军团菌;1份弗朗西斯菌检测HEX阳性(假阳性),占实际培养分离的1/26.结论 单一及双重荧光定量PCR法特异、快速、敏感,一次同时检测嗜肺与非嗜肺军团菌,满足对空调和环境水样军团菌监测的要求.     

应用单一和双重荧光定量PCR法快速检测军团菌

目的 建立单一和双重荧光定量PCR方法分别和同时进行军团菌属及嗜肺军团菌的检测.方法 利用军团菌属16 S rRNA基因和嗜肺军团菌mip基因设计引物和探针,两条基因探针分别标记FAM和HEX,并将相关反应体系和条件进行优化.分别应用单一基因探针(单一荧光定量PCR)和双重基因探针(双重荧光定量PCR)对嗜肺军团菌、非嗜肺军团菌及非军团菌进行检测,并验证两种方法的特异度、敏感度.应用双重荧光定量PCR检测空调水样滤膜样品和DNA提取样品,比较两者结果的一致性.结果 针对军团菌属及嗜肺军团菌,应用荧光定量PCR,16 S rRNA基因和mip基因均能较好的检出,16S rRNA和mip的最低检出限分别为8和10个拷贝.经优化得到了最佳反应体系.单一荧光定量PCR方法所检的8株嗜肺军用菌及4株非嗜肺军团菌16 S rRNA基因均为阳性,嗜肺军团菌mip基因阳性,非嗜肺军团菌mip基因阴性.双重荧光定量PCR方法所检的23株嗜肺军团菌中有2株为假阴性,9株非嗜肺军团菌和非军团菌属中有1株为假阳性.49份空调水样滤膜直接检测和提取DNA后检测的结果一致,其中26份水样军团菌阳性,20份为嗜肺军团菌,6份为非嗜肺军团菌;1份弗朗西斯菌检测HEX阳性(假阳性),占实际培养分离的1/26.结论 单一及双重荧光定量PCR法特异、快速、敏感,一次同时检测嗜肺与非嗜肺军团菌,满足对空调和环境水样军团菌监测的要求.     

间变性大细胞淋巴瘤患儿EEF1G/ALK融合基因的鉴定与检测

目的 明确1例伴少见ALK融合基因的间变性大细胞淋巴瘤患儿的分子诊断,建立该融合基因的实时荧光定量PCR检测方法。方法 采用基于锚定多重PCR的二代测序技术,鉴定患儿腹股沟肿大淋巴结组织中ALK基因的伙伴基因。建立EEF1G/ALK融合基因实时荧光定量PCR方法,并对方法的重复性、灵敏度和特异性进行性能评价。在此基础上检测此患儿肿瘤组织、骨髓、外周血和脑脊液EEF1G/ALK融合基因的表达。结果 经二代测序鉴定该患儿肿瘤细胞EEF1G/ALK融合基因阳性,Sanger测序验证为EEF1G Exon 6和ALK Exon 20的融合。建立的实时荧光定量PCR方法最低定量限为40拷贝/体系;弱阳性标本和强阳性标本的批内精密度CV分别为0.52%和0.17%;批间精密度CV分别为1.32%和1.14%,重复性好;检测其他ALK融合阳性标本结果为阴性,特异性好。复发时送检的淋巴结穿刺组织和外周血的融合基因定量检测结果为138.92%和0.0039%,其余检测为阴性。结论 明确此例间变性大细胞淋巴瘤为少见EEF1G/ALK融合基因阳性。建立的EEF1G/ALK融合基因定量检测方法特异性、重复性好...     

念珠菌耐药机制的研究进展

近年来,随着侵袭性真菌感染危险因素的增多,如器官移植、中性粒细胞减少、长期应用糖皮质类固醇激素等,侵袭性真菌感染的发病率和死亡率不断上升。临床上侵袭性真菌感染仍以念珠菌属感染为主,其中白念珠菌感染占54％、光滑念珠菌占22％、近平滑念珠菌占6％、热带念珠菌占5．7％、克柔念珠菌占4．3％。随着抗真菌药物长期反复广泛应用,对抗真菌药物耐药的念珠菌也愈来愈多,     

Rapid detection and identification of Candida albicans and Torulopsis (Candida) glabrata in clinical specimens by species-specific nested PCR amplification of a cytochrome P-450 lanosterol-alpha-demethylase (L1A1) gene fragment.

PCR of a Candida albicans cytochrome P-450 lanosterol-alpha-demethylase (P450-L1A1) gene segment is a rapid and sensitive method of detection in clinical specimens. This enzyme is a target for azole antifungal action. In order to directly detect and identify the clinically most important species of Candida, we cloned and sequenced 1.3-kbp fragments of the cytochrome P450-L1A1 genes from Torulopsis (Candida) glabrata and from Candida krusei. These segments were compared with the published sequences from C. albicans and Candida tropicalis. Amplimers for gene sequences highly conserved throughout the fungal kingdom were first used; positive PCR results were obtained for C. albicans, T. glabrata, C. krusei, Candida parapsilosis, C. tropicalis, Cryptococcus neoformans, and Trichosporon beigelii DNA extracts. Primers were then selected for a highly variable region of the gene, allowing the species-specific detection from purified DNA of C. albicans, T. glabrata, C. krusei, and C. tropicalis. The assay sensitivity as tested for C. albicans in seeded clinical specimens such as blood, peritoneal fluid, or urine was 10 to 20 cells per 0.1 ml. Compared with results obtained by culture, the sensitivity, specificity, and efficiency of the species-specific nested PCR tested with 80 clinical specimens were 71, 95, and 83% for C. albicans and 100, 97, and 98% for T. glabrata, respectively.     

Multiplex real-time PCR targeting the RNase P RNA gene for detection and identification of Candida species in blood

We have developed a single-tube multiplex real-time PCR method for the detection of the eight most common Candida species causing septicemia: Candida albicans, C. dubliniensis, C. famata, C. glabrata, C. guilliermondii, C. krusei, C. parapsilosis, and C. tropicalis. The method developed targets the RNase P RNA gene RPR1. Sequences of this gene were determined for seven of the Candida species and showed surprisingly large sequence variation. C. glabrata was found to have a gene that was five times longer gene than those of the other species, and the nucleotide sequence similarity between C. krusei and C. albicans was as low as 55%. The multiplex PCR contained three probes that enabled the specific detection of C. albicans, C. glabrata, and C. krusei and a fourth probe that allowed the general detection of the remaining species. The method was able to detect 1 to 10 genome copies when the detection limit was tested repeatedly for the four species C. albicans, C. glabrata, C. krusei, and C. guilliermondii. No significant difference in the detection limit was seen when the multiplex format was compared with single-species PCR, i.e., two primers and one probe. The method detected eight clinically relevant Candida species and did not react with other tested non-Candida species or human DNA. The assay was applied to 20 blood samples from nine patients and showed a sensitivity similar to that of culture.     

热带假丝酵母菌实时荧光定量PCR检测方法的建立及临床应用初步评价

目的建立、优化快速检测临床标本中热带假丝酵母菌的实时荧光定量PCR方法 ,并对其临床应用进行初步评价。方法用自行设计的高效引物、探针检测5种临床标本中的热带假丝酵母菌,对该方法的敏感性、特异性进行评价,并与真菌培养法进行比较。结果检测临床标本中热带假丝酵母菌的灵敏度可达10 copies/ml,与人类基因组、细菌及其他真菌无交叉阳性反应。与真菌培养法的检测一致性好（Kappa值为0.916,P〈0.01）。结论实时荧光定量PCR检测热带假丝酵母菌灵敏度高、特异性强,可直接用于各种临床标本中热带假丝酵母菌的检测,而且可大大缩短报告时间,为临床诊断提供可靠依据。     

Rapid identification of commonly encountered Candida species directly from blood culture bottles

We report a rapid-cycle, real-time PCR method for identifying six Candida spp. directly from BACTEC blood culture bottles. Target sequences in the noncoding internal transcribed spacer regions of the rRNA operon were simultaneously amplified and interrogated with fluorescent probes to identify Candida albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, and C. lusitaniae; these account for 88% of the yeast species isolated from positive blood cultures in our laboratory. Any of the first four species can be identified in a single reaction using two fluorescent hybridization probe sets. The antifungal-resistant species C. krusei and C. lusitaniae are detected in a second reaction, also with two probe sets. The assay was validated with DNA extracted from BACTEC blood culture bottles positive for yeasts (n = 62) and was 100% concordant with culture identification based on biochemical and morphological features of C. albicans (n = 22), C. parapsilosis (n = 10), C. tropicalis (n = 1) C. glabrata (n = 22), C. krusei (n = 2), and C. lusitaniae (n = 1). No cross-reactivity was observed in blood culture samples growing yeasts other than the above-mentioned species (n = 4), in those growing bacteria (n = 12), or in the absence of microbial growth. Our assay allows rapid (相似文献   

Comparison of three differential media for the presumptive identification of yeasts

This study evaluated three differential media, CHROMagar Candida, BiGGY agar and Albicans ID2 agar, for the presumptive identification of yeast species. In total, 215 yeast isolates were included in the study. The sensitivity and specificity of CHROMagar Candida, BiGGY agar and Albicans ID2 agar for the detection of Candida albicans were 100% and 100%, 91% and 92.7%, and 99.2% and 92.7%, respectively. CHROMagar Candida was a reliable tool for the presumptive identification of C. albicans, Candida tropicalis, Candida krusei and Candida glabrata. Albicans ID2 agar was useful for the detection of C. albicans.     

Development of novel real-time PCR assays for detection and differentiation of eleven medically important Aspergillus and Candida species in clinical specimens

In the present study, novel real-time PCR assays targeting the fungal ITS2 region were developed for the detection and differentiation of medically important Aspergillus species (Aspergillus fumigatus, Aspergillus flavus, Aspergillus nidulans, Aspergillus niger, and Aspergillus terreus) and Candida species (Candida albicans, Candida dubliniensis, Candida glabrata, Candida krusei, Candida parapsilosis, and Candida tropicalis) using a LightCycler instrument. The combination of a group-specific and a universal primer with five Aspergillus or six Candida species-specific biprobes in one reaction mixture facilitated rapid screening and species differentiation by the characteristic peak melting temperatures of the biprobes. Both assays can be performed either as single assays or simultaneously in the same LightCycler run. The analytical sensitivity using pure cultures and EDTA-anticoagulated blood, cerebrospinal fluid (CSF), and tissue samples spiked with A. fumigatus and C. albicans cell suspensions was shown to be at least 1 CFU per PCR, corresponding to 5 to 10 CFU/ml blood and 10 CFU/200 microl CSF or 0.02 g tissue. To assess the clinical applicability, 26 respiratory samples, 4 tissue samples from the maxillary sinus, and 1 blood sample were retrospectively tested and real-time PCR results were compared with results from culture, histology, or a galactomannan enzyme-linked immunosorbent assay (ELISA). Twenty samples (64.5%) were both culture positive and positive by real-time PCR. Six samples (19.4%) showed no growth of fungi but were positive by real-time PCR. However, all of the tissue samples were positive by both PCR and histology. The blood sample showed no growth of Aspergillus, but aspergillosis was confirmed by positive galactomannan ELISA, histology, and PCR results. The remaining samples (16.1%) were culture and PCR negative; also, no other signs indicating fungal infection were observed. Our data suggest that the Aspergillus and Candida assays may be appropriate for use in clinical laboratories as simple and rapid screening tests for the most frequently encountered Aspergillus and Candida species and might become an important tool in the early diagnosis of fungal infections in the future.     

Evaluation of nested and real-time PCR assays in the diagnosis of candidaemia

Polymerase chain reaction (PCR) assays have a very low theoretical detection threshold and are therefore advocated for the diagnosis of fungaemia. However, their effectiveness in this respect remains to be assessed. This study compared real-time PCR ( Can -G) and nested PCR assays with blood culture for the diagnosis of Candida spp. bloodstream infections. A total of 200 clinical blood samples obtained from 110 patients at risk for developing a systemic fungal infection, hospitalized in the University Hospital of Sfax (Tunisia), were submitted to testing by culture, nested PCR and real-time PCR. Blood culture was positive in 36 patients. When compared with culture, the Can -G assay (81% sensitivity, 96% specificity) performed better than the nested PCR assay (86% sensitivity, 54% specificity). The real-time PCR assay, which avoids both the contamination hazard with amplicons that may cause false-positive results and the use of time-consuming post-PCR steps, appears more suitable than the nested PCR assay for the laboratory diagnosis of Candida spp. bloodstream infections. In this study, real-time PCR did not enhance the diagnostic sensitivity for Candida spp. bloodstream infections compared with conventional blood culture; however, it may lead to earlier implementation of an adequately targeted antifungal treatment.     

Evaluation of the new chromogenic medium Candida ID 2 for isolation and identification of Candida albicans and other medically important Candida species

The usefulness of Candida ID 2 (CAID2) reformulated medium (bioMérieux, France) has been compared with that of the former Candida ID (CAID; bioMérieux), Albicans ID 2 (ALB2; bioMérieux), and CHROMagar Candida (CAC; Chromagar, France) chromogenic media for the isolation and presumptive identification of clinically relevant yeasts. Three hundred forty-five stock strains from culture collections, and 103 fresh isolates from different clinical specimens were evaluated. CAID2 permitted differentiation based on colony color between Candida albicans (cobalt blue; sensitivity, 91.7%; specificity, 97.2%) and Candida dubliniensis (turquoise blue; sensitivity, 97.9%; specificity, 96.6%). Candida tropicalis gave distinguishable pink-bluish colonies in 97.4% of the strains in CAID2 (sensitivity, 97.4%; specificity, 100%); the same proportion was reached in CAC, where colonies were blue-gray (sensitivity, 97.4%; specificity, 98.7%). CAC and CAID2 showed 100% sensitivity values for the identification of Candida krusei. However, with CAID2, experience is required to differentiate the downy aspect of the white colonies of C. krusei from other white-colony-forming species. The new CAID2 medium is a good candidate to replace CAID and ALB2, and it compares well to CAC for culture and presumptive identification of clinically relevant Candida species. CAID2 showed better results than CAC in some aspects, such as quicker growth and color development of colonies from clinical specimens, detection of mixed cultures, and presumptive differentiation between C. albicans and C. dubliniensis.     

Detection of serum Candida antigens by enzyme-linked immunosorbent assay and a latex agglutination test with anti-Candida albicans and anti-Candida krusei antibodies.

Serum samples from 197 patients with and without candidiasis were assayed for Candida albicans mannan and Candida krusei mannan by an enzyme-linked immunosorbent assay (ELISA) and a latex agglutination test (LA) and for D-arabinitol by the enzymatic fluorometric method. Of the 43 patients positive for C. albicans mannan (> or = 0.2 ng/ml), 34 were infected with C. albicans and 9 were infected with Candida tropicalis. C. krusei mannan (> or = 0.3 ng/ml) was detected in 10 patients infected with Candida parapsilosis, 2 patients infected with Candida guilliermondii, and 1 patient infected with C. krusei. With both anti-C. albicans antibodies and anti-C. krusei antibodies, the sensitivities of ELISA and LA for detection of invasive candidiasis (58 patients) were 74 and 38%, respectively. No false-positive reactions were observed by the ELISA or the LA. The sensitivity and specificity of the D-arabinitol/creatinine ratio (> or = 1.5 mumol/mg) to invasive candidiasis were 50 and 91%, respectively. The ELISA with antibodies against both C. albicans and C. krusei may be useful in diagnosing invasive candidiasis caused by medically important Candida strains excluding Candida glabrata.     

Use of a monoclonal antibody in a dot immunobinding assay for detection of a circulating mannoprotein of Candida spp. in neutropenic patients with invasive candidiasis.

A dot immunobinding assay for the detection of a circulating mannoprotein (MP) antigen of Candida species in the sera of neutropenic patients in a hematological setting is described. The technique is based on the use of a monoclonal antibody which recognizes an oligomannoside epitope shared by distinct MP of pathogenic Candida species. The sensitivity of the assay for antigen detection in serum was between 2 and 5 ng/ml, and MPs from Candida albicans, Candida tropicalis, Torulopsis glabrata, and Candida parapsilosis, but not Candida krusei, could be detected. A retrospective analysis of sera from patients with proven invasive candidiasis versus sera from controls (Candida-colonized and noncolonized subjects) revealed that the novel assay has sufficient sensitivity, specificity, and predictive values to be of potential diagnostic significance.