Rapid, Accurate, and Low-Cost Stool Clostridium difficile Genetic Test
快速、准确、低成本的粪便艰难梭菌基因检测
基本信息
- 批准号:8715249
- 负责人:
- 金额:$ 30万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2014
- 资助国家:美国
- 起止时间:2014-04-15 至 2016-03-31
- 项目状态:已结题
- 来源:
- 关键词:AntibioticsAntigensBacteriaBindingBiological AssayBlindedCaringCellsCessation of lifeClinicalClostridium difficileContractsCytolysisCytotoxinDNADNA amplificationDetectionDiagnosisDiarrheaEnzyme ImmunoassayFecesFilmFluorescent ProbesFrequenciesGene AmplificationGenesGeneticGenetic screening methodGoalsGoldHealthcareHigh temperature of physical objectHospitalsHourIn SituInfectionIntestinesLabelLeadLinkMagnesiumMediatingMedicalMonitorNucleic Acid Amplification TestsNucleic AcidsNursing HomesOutpatientsPatientsPersonsPhasePhysiciansPolymerase Chain ReactionPrevalence StudyRecurrenceRelative (related person)RiskSamplingSensitivity and SpecificitySeveritiesSpecificityStressSurfaceSystemTestingTimeToxincold temperaturecombatcostgalactitol 2-dehydrogenaselead titanatemortalitypatient safetypoint of careprototypepublic health relevancereceptorsensorsolid solution
项目摘要
DESCRIPTION (provided by applicant): Clostridium difficile (CD) is a bacterium causing diarrhea and other intestinal problems linked to 14,000 annual deaths in the US. CDI is an antibiotic-associated infection as well as a health-care-associated infection. Current CDI diagnosis relies on CD toxins enzyme immunoassay (EIA) together with antigen (GDH) EIA. Although stool toxin EIA is specific and relatively fast (hours to 1 day), the sensitivity of toxinEIA is only 60%. Nucleic acid amplification test (NAAT) using quantitative real-time polymerase chain reaction (qPCR) or loop-mediated isothermal amplification (LAMP) to detect the toxin gene tcdB or tcdA is sensitive and specific. However, due to the emergence of hypervirulent strains, detecting tcdA or tcdB alone cannot tell the severity of CDI, and is not sufficient to hel physician make informed CDI treatment decisions. The 30-day mortality rate and recurrence rate for CDI caused by hypervirulent strains are higher and are linked to a third toxin, the binary
toxin which is encoded by cdtB and cdtA. All CDI involve tcdB but not tcdA. Therefore, if there is a genetic CD test that can detect tcdB and cdtB simultaneously in one test it would be possible to not only diagnose CDI but also determine the severity and recurrence risk of the CDI to help physicians make treatment decisions. (PbMg1/3Nb2/3O3)0.65(PbTiO3)0.35 (PMN-PT) piezoelectric plate sensor (PEPS) is a unique sensor developed by Shih and Shih, which uses relative resonance frequency shift, Df/f to detect binding of a target to the receptor on the PEPS surface. What is unique about PMN-PT PEPS is that detection Df/f is enhanced by 1000 times due to the unique polarization switching mechanism within the PMN-PT layer. As a result, PEPS has demonstrated sensitivity similar to PCR in DNA detection but without the need of DNA extraction and DNA amplification. The goal of the study is to develop a rapid, accurate, and low-cost CDI test using a 3- PEPS array--one PEPS to detect tcdB, the second PEPS to detect cdtB, and the third PEPS to serve as a negative control with high sensitivity and specificity to help diagnose CDI and assessing CDI severity and recurrence risk. Encouraging preliminary results of detecting tcdB in 20 blinded patient stools showed that PEPS correctly identified 11/11 qPCR positive samples and 7/9 qPCR negative samples within 40 min using simple electrical means, indicating the sensitivity of the PEPS test. In addition, PEPS is easily multiplexed and the simple electrical detection makes the test low-cost and thus can be widely available.
描述(由申请人提供):艰难梭菌(CD)是一种引起腹泻和其他肠道问题的细菌,在美国每年有14,000人死亡。CDI是一种与疟疾相关的感染,也是一种与卫生保健相关的感染。目前CDI的诊断依赖于CD毒素酶免疫测定(EIA)和抗原(GDH)EIA。虽然粪便毒素EIA具有特异性和相对快速性(小时至1天),但ToxinEIA的敏感性仅为60%。核酸扩增试验(NAAT)采用实时定量聚合酶链反应(qPCR)或环介导等温扩增(LAMP)检测毒素基因tcdB或tcdA,具有敏感性和特异性。然而,由于高毒力菌株的出现,单独检测tcdA或tcdB不能判断CDI的严重程度,也不足以帮助医生做出明智的CDI治疗决定。由高毒力菌株引起的CDI的30天死亡率和复发率更高,并且与第三种毒素二元毒素有关
由cdtB和cdtA编码的毒素。所有CDI都涉及tcdB,但不涉及tcdA。因此,如果有一种遗传CD测试可以在一个测试中同时检测tcdB和cdtB,那么不仅可以诊断CDI,还可以确定CDI的严重程度和复发风险,以帮助医生做出治疗决策。(PbMg1/3Nb2/3O3)0.65(PbTiO3)0.35(PMN-PT)压电板传感器(PEPS)是由Shih和Shih开发的独特传感器,其使用相对共振频率偏移Df/f来检测靶与PEPS表面上的受体的结合。PMN-PT PEPS的独特之处在于,由于PMN-PT层内独特的偏振切换机制,检测Df/f增强了1000倍。因此,PEPS在DNA检测中表现出类似于PCR的灵敏度,但不需要DNA提取和DNA扩增。该研究的目标是使用3-PEPS阵列开发快速,准确和低成本的CDI测试-一个PEPS检测tcdB,第二个PEPS检测cdtB,第三个PEPS作为阴性对照,具有高灵敏度和特异性,以帮助诊断CDI并评估CDI严重程度和复发风险。在20个盲态患者粪便中检测tcdB的令人鼓舞的初步结果表明,PEPS使用简单的电气手段在40分钟内正确识别了11/11个qPCR阳性样本和7/9个qPCR阴性样本,表明PEPS测试的灵敏度。此外,PEPS易于多路复用,并且简单的电检测使得测试成本低,因此可以广泛使用。
项目成果
期刊论文数量(8)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(1)
Specific in situ hepatitis B viral double mutation (HBVDM) detection in urine with 60 copies ml(-1) analytical sensitivity in a background of 250-fold wild type without DNA isolation and amplification.
- DOI:10.1039/c4an01885k
- 发表时间:2015-03-07
- 期刊:
- 影响因子:0
- 作者:Kirimli CE;Shih WH;Shih WY
- 通讯作者:Shih WY
DNA hybridization detection with 100 zM sensitivity using piezoelectric plate sensors with an improved noise-reduction algorithm.
- DOI:10.1039/c4an00215f
- 发表时间:2014-06-07
- 期刊:
- 影响因子:0
- 作者:Kirimli CE;Shih WH;Shih WY
- 通讯作者:Shih WY
A model study of 3-dimensional localization of breast tumors using piezoelectric fingers of different probe sizes.
使用不同探头尺寸的压电手指进行乳腺肿瘤三维定位的模型研究。
- DOI:10.1063/1.5054287
- 发表时间:2019
- 期刊:
- 影响因子:0
- 作者:Xu,Xin;Shih,Wei-Heng;Shih,WanY
- 通讯作者:Shih,WanY
In situ, amplification-free double-stranded mutation detection at 60 copies/ml with thousand-fold wild type in urine.
- DOI:10.1016/j.bios.2018.07.062
- 发表时间:2018-11-15
- 期刊:
- 影响因子:12.6
- 作者:Kirimli C;Lin S;Su YH;Shih WH;Shih WY
- 通讯作者:Shih WY
Amplification-free in situ KRAS point mutation detection at 60 copies per mL in urine in a background of 1000-fold wild type.
- DOI:10.1039/c5an02048d
- 发表时间:2016-02-21
- 期刊:
- 影响因子:0
- 作者:Kirimli CE;Shih WH;Shih WY
- 通讯作者:Shih WY
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Wan Y. Shih其他文献
Sodium doped CsPbI<sub>3</sub>-PMMA composite electrospun fibrous membranes for aqueous photocatalyst and LED
- DOI:
10.1016/j.mseb.2024.117835 - 发表时间:
2025-02-01 - 期刊:
- 影响因子:
- 作者:
Haohan Yang;Ke Li;Yuang Ji;Donghai Lin;Wan Y. Shih;Wei-Heng Shih - 通讯作者:
Wei-Heng Shih
Wan Y. Shih的其他文献
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{{ truncateString('Wan Y. Shih', 18)}}的其他基金
A Low-Cost Portable Multiplex Serum Molecular Test for HBV and HCV Detection
用于检测 HBV 和 HCV 的低成本便携式多重血清分子测试
- 批准号:
8981624 - 财政年份:2015
- 资助金额:
$ 30万 - 项目类别:
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- 批准号:2022J011295
- 批准年份:2022
- 资助金额:10.0 万元
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结核分枝杆菌持续感染期抗原(latency antigens)的重组BCG疫苗研究
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- 批准年份:2008
- 资助金额:19.0 万元
- 项目类别:青年科学基金项目
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