title: Development of a Novel Urine Alzheimer-Associated Neuronal Thread Protein ELISA Kit and Its Potential Use in the Diagnosis of Alzheimer's Disease.
abstract: BACKGROUND
  Alzheimer's disease (AD) is an age-related chronic degenerative disease that damages the nervous system. A noninvasive and simple method for early detection of AD is extremely important for the diagnosis and prognosis of AD. Thus, we aimed to develop an enzyme-linked immunosorbent assay (ELISA) kit to detect urine Alzheimer-associated neuronal thread protein (AD7C-NTP), and to evaluate its clinical value for the diagnosis of AD.
  
  
  METHODS
  Immunogenic AD7C-NTP peptide fragments were synthesized by the solid-phase method and used for immunizing mice or rabbits to generate anti-AD7C-NTP antibodies. The urine AD7C-NTP ELISA kit was then established; the generated mouse anti-AD7C-NTP antibody was used as a capture antibody, the biotin-labeled rabbit anti-AD7C-NTP antibody was used as a detection antibody, and avidin labeled by horseradish peroxidase was used as a substrate. The first morning urine specimens of 121 AD patients and 118 age-matched controls were collected, and the urine AD7C-NTP levels were detected by the above ELISA kit.
  
  
  RESULTS
  Mouse and rabbit anti-AD7C-NTP antibody ELISA titer was found to be 1:8,000 and 1:32,000, respectively. A single band with a relative molecular mass of 41 kDa was found in human brain specimens by Western blot assay, which was identified as AD7C-NTP antibody. The urine AD7C-NTP concentration of the AD patients was higher than that of the age-matched controls, the sensitivity was 89.3% and the specificity was 84.7%.
  
  
  CONCLUSIONS
  Our study demonstrated that our newly developed urine AD7C-NTP ELISA kit has suggested potential for diagnosing AD in a Chinese population, suggesting it may be a useful diagnostic kit for detecting early AD.
authors.affiliation: Central Laboratory, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Disease of Ministry of Education, Center of Alzheimer's Disease, Beijing Institute for Brain Disorders, Beijing, China.|Department of Geriatrics, Xuanwu Hospital, Capital Medical University, Beijing, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Ma
authors.affiliation: Central Laboratory, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Disease of Ministry of Education, Center of Alzheimer's Disease, Beijing Institute for Brain Disorders, Beijing, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Neurology, Xuanwu Hospital, Capital Medical University, Beijing, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Han
authors.affiliation: Central Laboratory, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Disease of Ministry of Education, Center of Alzheimer's Disease, Beijing Institute for Brain Disorders, Beijing, China.
authors.forename: Shuli
authors.initials: S
authors.lastname: Sheng
authors.affiliation: Shenzhen Anqun Biotech CO., Ltd., Shenzhen, China.
authors.forename: Jianan
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Central Laboratory, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Disease of Ministry of Education, Center of Alzheimer's Disease, Beijing Institute for Brain Disorders, Beijing, China.
authors.forename: Zhijuan
authors.initials: Z
authors.lastname: Ji
authors.affiliation: Central Laboratory, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Disease of Ministry of Education, Center of Alzheimer's Disease, Beijing Institute for Brain Disorders, Beijing, China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Neurology, Beijing Chaoyanmen Hospital, Beijing, China.
authors.forename: Ziqing
authors.initials: Z
authors.lastname: Cao
authors.affiliation: Department of Clinical Laboratory, Xuanwu Hospital, Capital Medical University, Beijing, China.
authors.forename: Peichang
authors.initials: P
authors.lastname: Wang
mesh_terms: D000544:Alzheimer Disease / Q000175:diagnosis* / Q000652:urine*; D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005625:Frontal Lobe / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D009419:Nerve Tissue Proteins / Q000652:urine*; D011817:Rabbits; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: C497612:AD7c-NTP protein, human; D009419:Nerve Tissue Proteins
keywords: Alzheimer disease; Alzheimer-associated neuronal thread protein; enzyme-linked immunosorbent assay; urine