OBJECTIVES: Development of a generally applicable sensitive hybridization-based assay devoid of any target amplification for the detection and identification of (pathogenic) bacterial and viral species. DESIGN AND METHODS: Using a sandwich hybridization format, the presence of a species-specific nucleic acid sequence is detected by means of Lateral Flow (LF) and Up-converting Phosphor Technology (UPT, a luminescent tracer). As a model, detection of the pathogen Streptococcus pneumoniae was investigated using a probe against the single-copy lytA gene. RESULTS: Detection of S. pneumoniae (in particular a 1200 p lytA sequences) required less than 1 ng of genomic DNA (approximate size 2.2 Mb). Hybridization and detection were performed in a complex background containing 10 microg fish sperm DNA. CONCLUSIONS: The results indicate the possibility to detect nucleic acid targets in nonamplified DNA samples using easy, inexpensive, amplification-free hybridization-based assays and the ultra sensitive UPT reporters. Employment of UPT allows to by-pass target amplification and therefore brings genetic-based testing a step closer to the point-of-care environment. Detection of S. pneumoniae with only 1 ng of DNA indicates a potential for applications in the field of infectious diseases.
OBJECTIVES: Development of a generally applicable sensitive hybridization-based assay devoid of any target amplification for the detection and identification of (pathogenic) bacterial and viral species. DESIGN AND METHODS: Using a sandwich hybridization format, the presence of a species-specific nucleic acid sequence is detected by means of Lateral Flow (LF) and Up-converting Phosphor Technology (UPT, a luminescent tracer). As a model, detection of the pathogen Streptococcus pneumoniae was investigated using a probe against the single-copy lytA gene. RESULTS: Detection of S. pneumoniae (in particular a 1200 p lytA sequences) required less than 1 ng of genomic DNA (approximate size 2.2 Mb). Hybridization and detection were performed in a complex background containing 10 microg fish sperm DNA. CONCLUSIONS: The results indicate the possibility to detect nucleic acid targets in nonamplified DNA samples using easy, inexpensive, amplification-free hybridization-based assays and the ultra sensitive UPT reporters. Employment of UPT allows to by-pass target amplification and therefore brings genetic-based testing a step closer to the point-of-care environment. Detection of S. pneumoniae with only 1 ng of DNA indicates a potential for applications in the field of infectious diseases.
Authors: Paul L A M Corstjens; Lisette van Lieshout; Michel Zuiderwijk; Dieuwke Kornelis; Hans J Tanke; Andre M Deelder; Govert J van Dam Journal: J Clin Microbiol Date: 2007-10-17 Impact factor: 5.948
Authors: Paul L A M Corstjens; Michel Zuiderwijk; Hans J Tanke; Jolien J van der Ploeg-van Schip; Tom H M Ottenhoff; Annemiek Geluk Journal: Clin Biochem Date: 2008-01-03 Impact factor: 3.281
Authors: Paul L A M Corstjens; Anouk van Hooij; Elisa M Tjon Kon Fat; Susan J F van den Eeden; Louis Wilson; Annemieke Geluk Journal: Clin Vaccine Immunol Date: 2016-06-06
Authors: Paul L A M Corstjens; Claudia J de Dood; Jolien J van der Ploeg-van Schip; Karien C Wiesmeijer; Terhi Riuttamäki; Krista E van Meijgaarden; John S Spencer; Hans J Tanke; Tom H M Ottenhoff; Annemieke Geluk Journal: Clin Biochem Date: 2011-07-06 Impact factor: 3.281
Authors: G Ajithkumar; Benjamin Yoo; Dara E Goral; Peter J Hornsby; Ai-Ling Lin; Uma Ladiwala; Vinayak P Dravid; Dhiraj K Sardar Journal: J Mater Chem A Mater Date: 2013-03-21
Authors: Changchun Liu; Xianbo Qiu; Serge Ongagna; Dafeng Chen; Zongyuan Chen; William R Abrams; Daniel Malamud; Paul L A M Corstjens; Haim H Bau Journal: Lab Chip Date: 2008-12-05 Impact factor: 6.799