公开(公告)号：US08021846B2

公开(公告)日：2011-09-20

申请号：US12381492

申请日：2009-03-12

申请人： Scott E. Gygax ,  John-Paul Vermitsky ,  Sean G. Chadwick ,  Matthew J. Self ,  Eli Mordechai ,  Martin E. Adelson

发明人： Scott E. Gygax ,  John-Paul Vermitsky ,  Sean G. Chadwick ,  Matthew J. Self ,  Eli Mordechai ,  Martin E. Adelson

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6895 ,  C12Q2600/158 ,  C12Q2600/16

摘要： There is disclosed a method for determining azole resistance in Candida glabrata. A biological sample containing Candida glabrata is obtained and a normalized mRNA level of CDR1 gene is determined using qRT-PCR. Using a microbroth dilution assay conducted at azole concentrations of about 2-8 μg/mL, a susceptible isolate of Candida glabrata is obtained. A qRT-PCR assay is employed on the susceptible isolate and an average mRNA level of CDR1 is obtained. A fold-change value for CDR1 is obtained by comparing the CDR1 mRNA level of the biological sample with that of the average mRNA level. A ≧2-fold change value is indicative of an azole resistance in Candida glabrata. The present method provides a qRT-PCR assay for azole resistance that has a sensitivity of ≧90% and a specificity of ≧90%.

摘要翻译： 公开了一种测定念珠菌的唑类抗性的方法。 获得含有念珠菌的生物样品，并使用qRT-PCR测定CDR1基因的归一化mRNA水平。 使用在约2-8μg/ mL的唑浓度下进行的微量稀释测定，获得了易裂片假丝酵母（Candida glabrata）的分离物。 在易感性分离株上使用qRT-PCR测定，获得CDR1的平均mRNA水平。 通过将生物样品的CDR1 mRNA水平与平均mRNA水平的CDR1 mRNA水平进行比较，获得CDR1的倍数值。 A≥2倍变化值表示光滑念珠菌中的唑类抗性。 本方法提供具有≥90％的灵敏度和≥90％的特异性的唑抗性的qRT-PCR测定。

公开(公告)号：US20090305274A1

公开(公告)日：2009-12-10

申请号：US12381492

申请日：2009-03-12

申请人： Scott E. Gygax ,  John-Paul Vermitsky ,  Sean G. Chadwick ,  Matthew J. Self ,  Eli Mordechai ,  Martin E. Adelson

发明人： Scott E. Gygax ,  John-Paul Vermitsky ,  Sean G. Chadwick ,  Matthew J. Self ,  Eli Mordechai ,  Martin E. Adelson

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6895 ,  C12Q2600/158 ,  C12Q2600/16

摘要： There is disclosed a method for determining azole resistance in Candida glabrata. A biological sample containing Candida glabrata is obtained and a normalized mRNA level of CDR1 gene is determined using qRT-PCR. Using a microbroth dilution assay conducted at azole concentrations of about 2-8 μg/mL, a susceptible isolate of Candida glabrata is obtained. A qRT-PCR assay is employed on the susceptible isolate and an average mRNA level of CDR1 is obtained. A fold-change value for CDR1 is obtained by comparing the CDR1 mRNA level of the biological sample with that of the average mRNA level. A ≧2-fold change value is indicative of an azole resistance in Candida glabrata. The present method provides a qRT-PCR assay for azole resistance that has a sensitivity of ≧90% and a specificity of ≧90%.

摘要翻译： 公开了一种测定念珠菌的唑类抗性的方法。 获得含有念珠菌的生物样品，并使用qRT-PCR测定CDR1基因的标准化mRNA水平。 使用在约2-8mug / mL的唑浓度下进行的微量级稀释测定法，获得了易裂片假丝酵母（Candida glabrata）的分离物。 在易感性分离株上使用qRT-PCR测定，获得CDR1的平均mRNA水平。 通过将生物样品的CDR1 mRNA水平与平均mRNA水平的CDR1 mRNA水平进行比较，获得CDR1的倍数值。 A> = 2倍变化值表示光滑念珠菌中的唑类抗性。 本方法提供灵敏度> = 90％，特异性> = 90％的唑类抗性的qRT-PCR测定。