title: Mitogen-activated protein kinase 5, a novel molecular marker for the identification and detection of Trypanozoon species.
abstract: Based on the sequence of mitogen-activated protein kinase 5 (TbMAPK5) gene, we have developed a specific PCR method which can delineate the species within the Trypanozoon subgenus from other parasites or host DNA conveniently. In view of further application in field studies, we performed loop-mediated isothermal amplification (LAMP) employing filter paper flecked with rodent blood. Our data showed that TbMAPK5-specific LAMP was sensitive enough to detect a very low parasitemia during the early stages of the infection and fluctuating parasitemia period, and is below the detection limit by microscope. The detection limit for the infected blood sample was 1000 trypanosomes/ml of blood. Based on these results, we consider that TbMAPK5 locus may be a useful target for LAMP diagnosis providing sensitivity and the potential for the genotyping/identification of Trypanozoon species.
authors.affiliation: Center for Parasitic Organisms, State Key Laboratory of Biocontrol, School of Life Sciences and Key Laboratory of Tropical Diseases and Control of the Ministry of Education, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510275, PR China.
authors.forename: Ming-Hui
authors.initials: MH
authors.lastname: Wang
authors.forename: Yan-Zi
authors.initials: YZ
authors.lastname: Wen
authors.forename: Ying
authors.initials: Y
authors.lastname: Wei
authors.forename: De-Hua
authors.initials: DH
authors.lastname: Lai
authors.forename: Zakaria
authors.initials: Z
authors.lastname: Bengaly
authors.forename: Marc
authors.initials: M
authors.lastname: Desquesnes
authors.forename: Zhao-Rong
authors.initials: ZR
authors.lastname: Lun
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004311:Double-Blind Method; D005819:Genetic Markers; D005838:Genotype; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics*; D051379:Mice; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D018512:Parasitemia; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D015800:Protozoan Proteins / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013997:Time Factors; D014345:Trypanosoma / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan; D005819:Genetic Markers; D047908:Intracellular Signaling Peptides and Proteins; D015800:Protozoan Proteins; C111049:MAP-kinase-activated kinase 5; D017346:Protein Serine-Threonine Kinases