title: Detection of Plasmodium knowlesi DNA in the urine and faeces of a Japanese macaque (Macaca fuscata) over the course of an experimentally induced infection.
abstract: BACKGROUND
  Diagnostic techniques based on PCR for the detection of Plasmodium DNA can be highly sensitive and specific. The vast majority of these techniques rely, however, on the invasive sampling of blood from infected hosts. There is, currently, considerable interest in the possibility of using body fluids other than blood as sources of parasite DNA for PCR diagnosis.
  
  
  METHODS
  Urine and faeces were obtained from a Plasmodium knowlesi infected-Japanese macaque (Macaca fuscata) over the course of an experimentally induced infection. P. knowlesi DNA (PkDNA) extracted from urine and faeces were monitored by nested PCR targeting the P. knowlesi specific cytochrome b (cytb) gene.
  
  
  RESULTS
  Urinary PkDNA was detected on day 2, but was not amplified using DNA templates extracted from the samples on day 4, day 5 and day 6. Subsequently, urinary PkDNA was detected from day 7 until day 11, and from day 20 until day 30. PkDNA in faeces was detected from day 7 until day 11, and from day 20 until day 37. Moreover, real-time quantitative PCR showed a remarkable increase in the amount of urinary PkDNA following anti-malarial treatment. This might have been due to the release of a large amount of PkDNA from the degraded parasites as a result of the anti-malarial treatment, leading to excretion of PkDNA in the urine.
  
  
  CONCLUSIONS
  The cytb-PCR system using urine and faecal samples is of potential use in molecular epidemiological surveys of malaria. In particular, monkey faecal samples could be useful for the detection of zoonotic primate malaria in its natural hosts.
authors.affiliation: Laboratory of Tropical Medicine and Parasitology, Dokkyo Medical University, Mibu, Tochigi 321-0293, Japan. skawai@dokkyomed.ac.jp.
authors.forename: Satoru
authors.initials: S
authors.lastname: Kawai
authors.forename: Megumi
authors.initials: M
authors.lastname: Sato
authors.forename: Naoko
authors.initials: N
authors.lastname: Kato-Hayashi
authors.forename: Hisashi
authors.initials: H
authors.lastname: Kishi
authors.forename: Michael A
authors.initials: MA
authors.lastname: Huffman
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Maeno
authors.forename: Richard
authors.initials: R
authors.lastname: Culleton
authors.forename: Shusuke
authors.initials: S
authors.lastname: Nakazawa
mesh_terms: D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis / Q000652:urine*; D004195:Disease Models, Animal; D005243:Feces / Q000737:chemistry*; D005260:Female; D008251:Macaca / Q000652:urine*; D008288:Malaria / Q000378:metabolism / Q000469:parasitology* / Q000503:physiopathology / Q000652:urine*; D008853:Microscopy; D025202:Molecular Diagnostic Techniques; D010274:Parasitology; D016790:Plasmodium knowlesi / Q000235:genetics*; D016133:Polymerase Chain Reaction
chemical_list: D016054:DNA, Protozoan