التفاصيل البيبلوغرافية
| العنوان: |
VHH-phage based competitive real-time immuno-PCR for ultrasensitive detection of ochratoxin A in cereal ; Anal Chem |
| المساهمون: |
Liu, Xing, Xu, Yang, Xiong, Yong-hua, Tu, Zhui, Li, Yan-ping, He, Zhen-yun, Qiu, Yu-lou, Fu, Jin-heng, Gee, Shirley J., Hammock, Bruce D. |
| المصدر: |
Anal Chem. 86(15):7471-7477. |
| المجموعة: |
CDC Stacks (Centers for Disease Control and Prevention) |
| مصطلحات موضوعية: |
Article, VHH, ochratoxin A, real-time immuno-PCR, phage display |
| الوصف: |
Phage display-mediated immuno-polymerase chain reaction (PD-IPCR) is an ultrasensitive detection technology that combines the advantages of immuno-PCR and phage display. The phage particle, which displayed antibody fragments including single-chain fragment variable (scFv), variable domain of heavy-chain antibodies (VHH), and antigen-binding fragment (Fab) on the surface can be directly used in IPCR, supplying both the detection antibody and deoxyribonucleic acid (DNA) template. In this work, we used ochratoxin A (OTA) as a model system to study the capacity of PD-IPCR in the detection of toxic small molecular weight compounds, especially mycotoxins. An alpaca-derived VHH library was constructed and subjected to four cycles of panning. In total, 16 clones with four unique sequences were selected by competitive binding with OTA. The clone VHH-28 resulted in the lowest 50% inhibitory concentration of 0.31 ng/mL in the phage enzyme-linked immunosorbent assay (ELISA) and was selected to develop the VHH phage-based real-time immuno-PCR (RT-IPCR). The detection limit of the VHH phage-based RT-IPCR was 3.7 pg/L, with a linear range of 0.01-1000 pg/mL. This method was compared with conventional ELISA, and validation results indicated the reliability of VHH phage-based RT-IPCR in the detection of OTA in cereal samples. This study provides a new idea for the ultrasensitive detection of mycotoxins and other toxic small molecular weight compounds. ; 2U50OH007550/OH/NIOSH CDC HHS/United States ; P42 ES004699/ES/NIEHS NIH HHS/United States ; P42ES04699/ES/NIEHS NIH HHS/United States ; 2015-08-05T00:00:00Z ; 24992514 ; PMC4306448 |
| نوع الوثيقة: |
other/unknown material |
| اللغة: |
unknown |
| العلاقة: |
http://stacks.cdc.gov/view/cdc/30262Test/ |
| الإتاحة: |
http://stacks.cdc.gov/view/cdc/30262Test/ |
| رقم الانضمام: |
edsbas.15188262 |
| قاعدة البيانات: |
BASE |
