title: Early detection of novel Leishmania species DNA in the saliva of two HIV-infected patients.
abstract: BACKGROUND
  Leishmaniasis caused by two new species of Leishmania; L. siamensis and L. martiniquensis have been recently described in Thailand. The disease has mainly been documented in AIDS patients from southern Thailand. In this study, polymerase chain reaction (PCR) was used to determine HIV-Leishmania co-infection in southern Thailand.
  
  
  METHODS
  One ml of saliva and 3 ml of EDTA blood were collected from HIV-infected patients for PCR detection of Leishmania DNA, cloning and sequencing. The positive PCR samples were then cultured on Schneider's insect medium.
  
  
  RESULTS
  Three out of 316 saliva samples collected from HIV-infected patients were found to be positive for Leishmania DNA (0.95%). Among the positive samples, one patient was observed with disseminated cutaneous lesions and also tested positive via saliva, whole blood and buffy coat in PCR. The second case presenting with nodular lesions also gave a positive saliva test via PCR two months prior to buffy coat. This diagnosis was confirmed by microscopic examination and a culture of biopsy samples from a nodule. The last case was an asymptomatic Leishmania infection which tested PCR positive only in saliva with a consecutive sample collection conducted for three months.
  
  
  CONCLUSIONS
  The prevalence of Leishmania infection in HIV infected patients within this study is 0.95%. Leishmania DNA was detected in saliva by PCR prior to blood and buffy coat of two HIV infected patients. Early detection of Leishmania DNA in saliva would be beneficial for the follow up of asymptomatic Leishmania infected patients, the early treatment of leishmaniasis and for surveillance survey purpose. However, full evaluation of sensitivity and specificity of this technique with a large cohort of patients is required before deployment.
authors.affiliation: Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.|Excellence Center for Emerging Infectious Disease, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, 10330, Thailand.
authors.forename: Padet
authors.initials: P
authors.lastname: Siriyasatien
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, Faculty of Medicine, Prince of Songkla University, Songkhla, 90110, Thailand.
authors.forename: Sarunyou
authors.initials: S
authors.lastname: Chusri
authors.affiliation: Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Kanyarat
authors.initials: K
authors.lastname: Kraivichian
authors.affiliation: Department of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand.
authors.forename: Narissara
authors.initials: N
authors.lastname: Jariyapan
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, Faculty of Medicine, Prince of Songkla University, Songkhla, 90110, Thailand.
authors.forename: Thanaporn
authors.initials: T
authors.lastname: Hortiwakul
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, Faculty of Medicine, Prince of Songkla University, Songkhla, 90110, Thailand.
authors.forename: Khachornsakdi
authors.initials: K
authors.lastname: Silpapojakul
authors.affiliation: School of Life Sciences, University of Liverpool, Liverpool, UK.
authors.forename: Adam M
authors.initials: AM
authors.lastname: Pym
authors.affiliation: Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand. amphumee@gmail.com.
authors.forename: Atchara
authors.identifier: http://orcid.org/0000-0001-8243-7406
authors.initials: A
authors.lastname: Phumee
mesh_terms: D016054:DNA, Protozoan / Q000032:analysis*; D042241:Early Diagnosis; D015658:HIV Infections / Q000150:complications / Q000469:parasitology; D006801:Humans; D007896:Leishmaniasis / Q000175:diagnosis / Q000469:parasitology; D012463:Saliva / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan
keywords: HIV-infection; Leishmania; PCR; Saliva