title: Emergence of a new focus of Plasmodium malariae in forest villages of district Balaghat, Central India: implications for the diagnosis of malaria and its control.
abstract: OBJECTIVE
  During an epidemiological study (January-July 2012) on malaria in forest villages of Central India, Plasmodium malariae-like malaria parasites were observed in blood smears of fever cases. We aimed to confirm the presence of P. malariae using molecular tools i.e. species-specific nested polymerase chain reaction (PCR) and DNA sequencing.
  
  
  METHODS
  All fever cases or cases with history of fever in 25 villages of Balaghat district were screened for malaria parasite using bivalent rapid diagnostic test and microscopy after obtaining written informed consent. Nested PCR was employed on microscopically suspected P. malariae cases. DNA sequences in the target region for PCR diagnosis were analysed for all the suspected cases of P. malariae.
  
  
  RESULTS
  Among the 22 microscopy suspected P. malariae cases, nested PCR confirmed the identity of P. malariae in 19 cases. Among these 14 were mono P. malariae infections, three were mixed infection of P. malariae with Plasmodium falciparum and two were mixed infection of P. malariae with Plasmodium vivax. Clinically P. malariae subjects generally presented with fever and headache. However, the typical 3-day pattern of quantum malaria was not observed. The parasite density of P. malariae was significantly lower than that of P. vivax and P. falciparum.
  
  
  CONCLUSIONS
  Plasmodium malariae may have been in existence in forest villages of central India but escaped identification due to its close resemblance to P. vivax. The results re-affirm the importance of molecular methods of testing on routine basis for efficacious control strategies against malaria.
authors.affiliation: Regional Medical Research Centre for Tribals, Jabalpur, Madhya Pradesh, India.
authors.forename: Praveen K
authors.initials: PK
authors.lastname: Bharti
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Chand
authors.forename: Mrigendra P
authors.initials: MP
authors.lastname: Singh
authors.forename: Sweta
authors.initials: S
authors.lastname: Mishra
authors.forename: Man M
authors.initials: MM
authors.lastname: Shukla
authors.forename: Rajesh
authors.initials: R
authors.lastname: Singh
authors.forename: Neeru
authors.initials: N
authors.lastname: Singh
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D015373:Bacterial Typing Techniques; D002648:Child; D002675:Child, Preschool; D016054:DNA, Protozoan; D005334:Fever / Q000097:blood / Q000209:etiology / Q000469:parasitology*; D006801:Humans; D007194:India; D007223:Infant; D017053:Infection Control / Q000379:methods*; D008288:Malaria / Q000097:blood / Q000175:diagnosis / Q000469:parasitology* / Q000517:prevention & control; D008853:Microscopy; D008875:Middle Aged; D010963:Plasmodium falciparum / Q000235:genetics; D010965:Plasmodium malariae / Q000235:genetics; D010966:Plasmodium vivax / Q000235:genetics; D016133:Polymerase Chain Reaction; D012424:Rural Population; D014197:Trees; D055815:Young Adult
chemical_list: D016054:DNA, Protozoan