title: Detection of chloroquine and artemisinin resistance molecular markers in Plasmodium falciparum: A hospital based study.
abstract: BACKGROUND
  Emergence of chloroquine (CQ) resistance in Plasmodium falciparum has increased the morbidity and mortality of falciparum malaria worldwide. Artemisinin-based combination therapies are now recommended by the World Health Organization as the first line treatment for falciparum malaria. Numerous molecular markers have been implicated in the CQ and artemisinin resistance.
  
  
  METHODS
  A total of 26 confirmed cases of falciparum malaria (by giemsa stained thick and thin smear, quantitative buffy coat, immunochromatographic test, or polymerase chain reaction [PCR]) were included in the study. About 5 ml of ethylenediaminetetraacetic acid blood sample was collected and stored at -20°C till use. Plasmodium DNA was extracted using QIAamp whole blood DNA extraction kit. PCR was done to amplify pfcrt, pfmdr1, pfserca, and pfmrp1 genes and the amplicons obtained were sequenced by Macrogen, Inc., Korea. Single nucleotide polymorphism (SNP) analysis was done using Bio-Edit Sequence Alignment Editor.
  
  
  RESULTS
  Out of the four genes targeted, we noted a SNP in the pfcrt gene alone. This SNP (G > T) was noted in the 658(th) position of the gene, which was seen in 13 patients. The pfmdr1 and pfserca genes were present in 9 and 14 patients respectively. But we could not find any SNPs in these genes. This SNP in pfcrt gene was not significantly associated with any adverse outcome and neither altered disease progression.
  
  
  CONCLUSIONS
  Presence of a single SNP may not be associated with any adverse clinical outcome. As the sample size was small, we may have not been able to detect any other known or unknown polymorphisms.
authors.affiliation: Department of Microbiology, Jawaharlal Nehru Institute of Postgraduate Medical Education and Research, Puducherry, India.
authors.forename: S
authors.initials: S
authors.lastname: Ramani
authors.affiliation: Department of Microbiology, Jawaharlal Nehru Institute of Postgraduate Medical Education and Research, Puducherry, India.
authors.forename: Subhash Chandra
authors.initials: SC
authors.lastname: Parija
authors.affiliation: Department of Microbiology, Jawaharlal Nehru Institute of Postgraduate Medical Education and Research, Puducherry, India.
authors.forename: Jharna
authors.initials: J
authors.lastname: Mandal
authors.affiliation: Department of Medicine, Jawaharlal Nehru Institute of Postgraduate Medical Education and Research, Puducherry, India.
authors.forename: Abdoul
authors.initials: A
authors.lastname: Hamide
authors.affiliation: Department of Paediatrics, Jawaharlal Nehru Institute of Postgraduate Medical Education and Research, Puducherry, India.
authors.forename: Vishnu
authors.initials: V
authors.lastname: Bhat
keywords: Chloroquine; quantitative buffy coat; single nucleotide polymorphisms