title: Enzymatic reporting of peste des petits ruminants virus genes ligating two specific probes on nanoparticles.
abstract: An alternative strategy for the detection of nucleic acid derived from peste des petits ruminants virus was developed omitting amplification. The assay is based on two probes complementary to the target sequences, one conjugated to magnetic microparticles the second to gold nanoparticles labeled with horseradish peroxidase. In the presence of target gene the two particles ligate via the probes and the complex can be magnetically separated. Applying substrate and chromogen a color reaction results for a positive case. Under optimized conditions, the approach had a linear detection range from 10 fM to 1 μM for ssDNA corresponding to an RNA low detection limit of 17.6 ng/μl. The quick performance (45 min) and not requiring expensive instrumentations offer a new way of detecting nucleic acids for the clinical diagnosis in our case for peste des petits ruminants virus.
authors.affiliation: State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Chunai
authors.initials: C
authors.lastname: Tao
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Huaxin
authors.initials: H
authors.lastname: Huang
mesh_terms: D011088:DNA Ligases / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000032:analysis; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D053758:Nanoparticles; D015345:Oligonucleotide Probes / Q000378:metabolism; D018107:Peste-des-petits-ruminants virus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D015345:Oligonucleotide Probes; D012367:RNA, Viral; D006735:Horseradish Peroxidase; D011088:DNA Ligases