BACKGROUND: In the TEDDY (The Environmental Determinants of Diabetes in the Young) study patient eligibility is based on the presence of some selected type 1 diabetes risk-associated human leukocyte antigen DR-DQ genotypes. A practical screening strategy was needed with efficient exclusion of ineligible patients at an early stage. Also, a simple, low-cost, and fast screening system was essential for the primary step of the risk assessment including thousands of samples. METHODS: A homogeneous genotyping system utilizing an asymmetric polymerase chain reaction (PCR) and subsequent hybridization of allele-specific probes was designed to be used as the first screening step. This assay was combined with methods further elucidating the genetic risk of type 1 diabetes to screen for high-risk individuals. RESULTS: The homogeneous assay platform allows the typing of hundreds of samples within one working day. The costs of the assay are minimal, and the reduction in hands-on time provides considerable improvements compared to the heterogeneous genotyping methods comprising separate PCR and hybridization steps. The primary selection criteria used in the first step proved to be efficient since the numbers of samples typed in subsequent stages were markedly reduced. CONCLUSIONS: The presented assay system provides a practical approach to the rapid screening of thousands of samples at low cost, a general starting point for large-scale screening studies.
BACKGROUND: In the TEDDY (The Environmental Determinants of Diabetes in the Young) study patient eligibility is based on the presence of some selected type 1 diabetes risk-associated human leukocyte antigen DR-DQ genotypes. A practical screening strategy was needed with efficient exclusion of ineligible patients at an early stage. Also, a simple, low-cost, and fast screening system was essential for the primary step of the risk assessment including thousands of samples. METHODS: A homogeneous genotyping system utilizing an asymmetric polymerase chain reaction (PCR) and subsequent hybridization of allele-specific probes was designed to be used as the first screening step. This assay was combined with methods further elucidating the genetic risk of type 1 diabetes to screen for high-risk individuals. RESULTS: The homogeneous assay platform allows the typing of hundreds of samples within one working day. The costs of the assay are minimal, and the reduction in hands-on time provides considerable improvements compared to the heterogeneous genotyping methods comprising separate PCR and hybridization steps. The primary selection criteria used in the first step proved to be efficient since the numbers of samples typed in subsequent stages were markedly reduced. CONCLUSIONS: The presented assay system provides a practical approach to the rapid screening of thousands of samples at low cost, a general starting point for large-scale screening studies.
Authors: Paul Dantonio; Nancy Meredith-Molloy; William A Hagopian; Jin Xiong She; Beena Akolkar; Suzanne K Cordovado; Miyono Hendrix; L Omar Henderson; W Harry Hannon; Robert F Vogt Journal: J Diabetes Sci Technol Date: 2010-07-01
Authors: B Jonsdottir; C Andersson; A Carlsson; A Delli; G Forsander; J Ludvigsson; C Marcus; U Samuelsson; E Ortqvist; A Lernmark; S-A Ivarsson; H Elding Larsson Journal: Diabetologia Date: 2013-05-12 Impact factor: 10.122
Authors: Ahmed J Delli; Fariba Vaziri-Sani; Bengt Lindblad; Helena Elding-Larsson; Annelie Carlsson; Gun Forsander; Sten A Ivarsson; Johnny Ludvigsson; Ingrid Kockum; Claude Marcus; Ulf Samuelsson; Eva Örtqvist; Leif Groop; George P Bondinas; George K Papadopoulos; Åke Lernmark Journal: Diabetes Date: 2012-07-10 Impact factor: 9.461
Authors: Andrea K Steck; Helena Elding Larsson; Xiang Liu; Riitta Veijola; Jorma Toppari; William A Hagopian; Michael J Haller; Simi Ahmed; Beena Akolkar; Åke Lernmark; Marian J Rewers; Jeffrey P Krischer Journal: Pediatr Diabetes Date: 2017-01-27 Impact factor: 3.409
Authors: A Carlsson; I Kockum; B Lindblad; L Engleson; A Nilsson; G Forsander; A-K Karlsson; A Kernell; J Ludvigsson; C Marcus; I Zachrisson; S-A Ivarsson; A Lernmark Journal: Int J Obes (Lond) Date: 2011-06-28 Impact factor: 5.095