OBJECTIVE: To construct a lipL32/1-lipL21-OmpL1/2 fusion gene and its prokaryotic expression system, and to establish an enzyme-linked immunosorbent assay (ELISA) using the rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for sensitive and specific detection of IgM in the serum of patients with leptospirosis. METHODS: lipL32/1-lipL21-OmpL1/2 fusion genes were constructed using a primer-linking PCR. The target recombinant protein antigens, rLipL32/1, rLipL21, rOmpL1/2 and rLipL32/1-LipL21-OmpL1/2, were expressed and the purified antigens were then immobilized to the surface of microplate wells for ELISA-based detection of IgM in the sera of leptospirosis patients. RESULTS: Of 493 acute leptospirosis patients, 95.7% and 97.8% were positive by rLipL32/1-LipL21- OmpL1/2-IgM-ELISA using different serum dilutions, which was higher than the rLipL32/1-IgM-ELISA (93.1% and 90.3%), rLipL21-IgM-ELISA (90.3% and 87.0%), and rOmpL1-IgM-ELISA (85.6% and 81.1%) (P<0.01). All IgM-ELISAs tested negative against 56 non-leptospirosis patients with typhoid fever, hemorrhagic fever or dengue fever. CONCLUSION: Trigeminal fusion antigen increases ELISA sensitivity and the rLipL32/1-LipL21-OmpL1/2- IgM-ELISA is a sensitive and specific serological diagnostic method for clinical leptospirosis.
OBJECTIVE: To construct a lipL32/1-lipL21-OmpL1/2 fusion gene and its prokaryotic expression system, and to establish an enzyme-linked immunosorbent assay (ELISA) using the rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for sensitive and specific detection of IgM in the serum of patients with leptospirosis. METHODS: lipL32/1-lipL21-OmpL1/2 fusion genes were constructed using a primer-linking PCR. The target recombinant protein antigens, rLipL32/1, rLipL21, rOmpL1/2 and rLipL32/1-LipL21-OmpL1/2, were expressed and the purified antigens were then immobilized to the surface of microplate wells for ELISA-based detection of IgM in the sera of leptospirosispatients. RESULTS: Of 493 acute leptospirosispatients, 95.7% and 97.8% were positive by rLipL32/1-LipL21- OmpL1/2-IgM-ELISA using different serum dilutions, which was higher than the rLipL32/1-IgM-ELISA (93.1% and 90.3%), rLipL21-IgM-ELISA (90.3% and 87.0%), and rOmpL1-IgM-ELISA (85.6% and 81.1%) (P<0.01). All IgM-ELISAs tested negative against 56 non-leptospirosispatients with typhoid fever, hemorrhagic fever or dengue fever. CONCLUSION: Trigeminal fusion antigen increases ELISA sensitivity and the rLipL32/1-LipL21-OmpL1/2- IgM-ELISA is a sensitive and specific serological diagnostic method for clinical leptospirosis.
Authors: Luis G V Fernandes; Monica L Vieira; Karin Kirchgatter; Ivy J Alves; Zenaide M de Morais; Silvio A Vasconcellos; Eliete C Romero; Ana L T O Nascimento Journal: Infect Immun Date: 2012-07-16 Impact factor: 3.441
Authors: Huan Wang; Yifei Wu; David M Ojcius; X Frank Yang; Chenglin Zhang; Shibiao Ding; Xu'ai Lin; Jie Yan Journal: PLoS One Date: 2012-08-01 Impact factor: 3.240
Authors: Amy M Dechet; Michele Parsons; Madan Rambaran; Pheona Mohamed-Rambaran; Anita Florendo-Cumbermack; Shamdeo Persaud; Shirematee Baboolal; Mary D Ari; Sean V Shadomy; Sherif R Zaki; Christopher D Paddock; Thomas A Clark; Lazenia Harris; Douglas Lyon; Eric D Mintz Journal: PLoS One Date: 2012-07-09 Impact factor: 3.240
Authors: Carolina Lessa-Aquino; Camila Borges Rodrigues; Jozelyn Pablo; Rie Sasaki; Algis Jasinskas; Li Liang; Elsio A Wunder; Guilherme S Ribeiro; Adam Vigil; Ricardo Galler; Douglas Molina; Xiaowu Liang; Mitermayer G Reis; Albert I Ko; Marco Alberto Medeiros; Philip L Felgner Journal: PLoS Negl Trop Dis Date: 2013-10-17
Authors: Safar Ali Alizadeh; Seyyed Saeed Eshraghi; Mohammad Reza Pourmand; Taghi Naserpour; Gholamreza Abdollahpour; Abbas Rahimiforoshani; Reza Najafipour Journal: Iran Red Crescent Med J Date: 2014-03-05 Impact factor: 0.611