| Objective: To develop the LAMP for detection of herpes simple virus type 1, herpes simple virus type 2, human papillomavirus type 6 and human papillomavirus type 11,respectively.Methods: Four primers were designed to amplificate target gene of herpes simple virus type 1, herpes simple virus type 2, human papillomavirus type 6 and human papillomavirus type 11 by PrimerExplorer V4, respectively. Virus genomic DNA was extracted by Kit. Through optimizing the amplification conditions, we have established specifical LAMP for the correspondence pathogens. Target DNA amplified under isothermal conditions (60～65°C) for 60 min, then, results were judged by naked eyes and electrophoretic analysis. The products of LAMP were confirmed by digestion using restriction enzymes. To evaluate the specificity of the LAMP assay, other pathogens were used as controls and detected by LAMP . To compare the sensitivity between LAMP and PCR, target pathogens DNA were 10-fold serially diluted and were amplified by LAMP and PCR, respectively.ResuLts: After LAMP reaction, ladder patterns unique to the LAMP assay were observed with four LAMP for the target pathogens. Both naked eyes and electrophoretic analysis were able to detect the products in the LAMP assay. Amplification was not observed when negative controls were tested. The specificity of LAMP products for the target pathogens were also confirmed by restriction enzymes. The result demonstrate that LAMP with equal sensitivity to conventional PCR assay, and the detection limit of LAMP assay for HSV-1, HSV-2, HPV-6 and HPV-11 DNA was 1 pg/μL,10 pg/μL,1 pg/μL,1 pg/μL, respectively.Conclusion: LAMP assay for detection herpes simple virus type 1, herpes simple virus type 2, human papillomavirus type 6, human papillomavirus type 11 were established and the assay is rapid, specific and sensitive.
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