SUN AiHua; WANG Yuan; DU Peng; WU ShengLing; YAN Jie

doi:10.3967/0895-3988.2011.03.013

doi: 10.3967/0895-3988.2011.03.013

基金项目: the National Science and Technology Key Program for Infectious Diseases of China(2008ZX10004-015)%the Natural Science Foundation of Zhejiang Medical College in China(2007XZA02)

A Sensitive and Specific IgM-ELISA for the Serological Diagnosis of Human Leptospirosis Using a rLipL32/1-LipL21-OmpL1/2 Fusion Protein

Zhejiang Medical College, Hangzhou 310053, Zhejiang, China;Department of Medical Microbiology and Parasitology, College of Medicine, Zhejiang University, Hangzhou 310058, Zhejiang, China
The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, Zhejiang, China
Zhejiang Medical College, Hangzhou 310053, Zhejiang, China
Department of Medical Microbiology and Parasitology, College of Medicine, Zhejiang University, Hangzhou 310058, Zhejiang, China

Funds: the National Science and Technology Key Program for Infectious Diseases of China(2008ZX10004-015)%the Natural Science Foundation of Zhejiang Medical College in China(2007XZA02)

摘要: Objective To construct a HpL32/1-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 HpL32/1-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-lgM-EUSA using different serum dilutions, which was higher than the rLipL32/1-lgM-ELISA (93.1% and 90.3%), rLipL21-lgM-ELISA (90.3% and 87.0%), and rOmpL1-lgM-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-EUSA is a sensitive and specific serological diagnostic method for clinical leptospirosis.
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Abstract: Objective To construct a HpL32/1-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 HpL32/1-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-lgM-EUSA using different serum dilutions, which was higher than the rLipL32/1-lgM-ELISA (93.1% and 90.3%), rLipL21-lgM-ELISA (90.3% and 87.0%), and rOmpL1-lgM-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-EUSA is a sensitive and specific serological diagnostic method for clinical leptospirosis.
- Leptospira /
- Outer membrane protein /
- Fusion antigen /
- Recombinant expression /
- IgM-ELISA

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doi: 10.3967/0895-3988.2011.03.013

A Sensitive and Specific IgM-ELISA for the Serological Diagnosis of Human Leptospirosis Using a rLipL32/1-LipL21-OmpL1/2 Fusion Protein

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A Sensitive and Specific IgM-ELISA for the Serological Diagnosis of Human Leptospirosis Using a rLipL32/1-LipL21-OmpL1/2 Fusion Protein

doi: 10.3967/0895-3988.2011.03.013

English Abstract

A Sensitive and Specific IgM-ELISA for the Serological Diagnosis of Human Leptospirosis Using a rLipL32/1-LipL21-OmpL1/2 Fusion Protein

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目录

doi: 10.3967/0895-3988.2011.03.013

A Sensitive and Specific IgM-ELISA for the Serological Diagnosis of Human Leptospirosis Using a rLipL32/1-LipL21-OmpL1/2 Fusion Protein

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出版历程

A Sensitive and Specific IgM-ELISA for the Serological Diagnosis of Human Leptospirosis Using a rLipL32/1-LipL21-OmpL1/2 Fusion Protein

doi: 10.3967/0895-3988.2011.03.013

English Abstract

A Sensitive and Specific IgM-ELISA for the Serological Diagnosis of Human Leptospirosis Using a rLipL32/1-LipL21-OmpL1/2 Fusion Protein

全文HTML

目录