公开(公告)号：US06077665A

公开(公告)日：2000-06-20

申请号：US833677

申请日：1997-04-08

申请人： Erica E. Weirich ,  Ronald L. Rabin ,  Yvonne Maldonado ,  William E. Benitz ,  Leonore A. Herzenberg ,  Leonard A. Herzenberg

发明人： Erica E. Weirich ,  Ronald L. Rabin ,  Yvonne Maldonado ,  William E. Benitz ,  Leonore A. Herzenberg ,  Leonard A. Herzenberg

IPC分类号： G01N33/53 ,  C12Q1/02 ,  G01N33/569 ,  C12Q1/68

CPC分类号： G01N33/56972 ,  G01N33/569 ,  G01N33/56988

摘要： A rapid assay for infection in immunodeficient patients such as neonates or immunocompromised patients (e.g. HIV or transplant patients) allows diagnosis at initial evaluation, such that antibiotic treatment and confinement to an intensive care unit can be avoided for uninfected patients. The assay can be used for detecting bacterial, viral, or fungal colonization of the blood stream, cerebrospinal fluid (CSF), or urinary tract. The method is particularly useful for sepsis diagnosis. Polymorphonuclear leukocyte (PMN, neutrophil) CD11b (Mac-1, CR3) levels are measured by flow cytometry or laser scanning microscopy in low volume (0.1 ml) whole blood samples. A dual-laser FACS identifies neutrophils by FITC-conjugated anti-CD15 fluorescent antibodies, and identifies surface neutrophil CD11b marked with PE-conjugated anti-CD11b antibodies. Spontaneous upregulation of CD11b is prevented by handling samples at 4.degree. C. or adding a stabilizing compound such as anti-CD14 antibody or adenosine to the samples. A kit comprises calibration markers such as CD11b-coated microspheres for calibrating the device used for analysis according to diagnosis threshold(s).

摘要翻译： 免疫缺陷患者如新生儿或免疫受损患者（例如HIV或移植患者）中的感染的快速测定允许在初始评估时进行诊断，从而可以避免对未感染患者的抗生素治疗和对重症监护病房的限制。 该测定法可用于检测血流，脑脊液（CSF）或尿道的细菌，病毒或真菌定植。 该方法对败血症诊断特别有用。 通过流式细胞术或激光扫描显微镜在低体积（0.1ml）全血样品中测量多形核白细胞（PMN，嗜中性粒细胞）CD11b（Mac-1，CR3）水平。 双激光FACS通过FITC缀合的抗CD15荧光抗体识别嗜中性粒细胞，并鉴定用PE缀合的抗CD11b抗体标记的表面嗜中性粒细胞CD11b。 通过在4℃处理样品或向样品中加入稳定化合物如抗-CD14抗体或腺苷来防止CD11b的自发上调。 试剂盒包括诸如CD11b涂覆的微球的校准标记，用于根据诊断阈值校准用于分析的装置。