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Background  Botswana is one of eight SADC countries targeting malaria elimination by 2018. Through spirited upscaling of control activities and passive surveillance, significant reductions in case incidence of  Plasmodium f alciparum (0.96 – 0.01) was achieved between 2008 and 2012. As part of the elimination campaign, active detection of asymptomatic  Plasmodium  species by a highly sensitive method was deemed necessary. This study was carried out to determine asymptomatic  Plasmodium  species carriage by nested PCR in the country, in 2012.    Method  A cross-sectional study involving 3924 apparently healthy participants were screened for  Plasmodium  species in 14 districts (5 endemic: Okavango, Ngami, Tutume, Boteti and Bobirwa; and 9 epidemic: North East, Francistown, Serowe-Palapye, Ghanzi, Kweneng West, Kweneng East, Kgatleng, South East, and Good Hope). Venous blood was taken from each participant for a nested PCR detection of  Plasmodium  species.    Results  The parasite rates of asymptomatic  Plasmodium  species detected were as follows:  Plasmodium falciparum , 0.16 %;  Plasmodium vivax , 4.66 %;  Plasmodium malariae , ( Pm ) 0.16 %;  Plasmodium ovale , 0 %, mixed infections ( P. falciparum  and  P. vivax ), 0.055 %; and ( P. vivax  and  P. malariae ), 0.027 %, (total: 5.062 %). The high proportion of asymptomatic reservoir of  P. vivax  was clustered in the East, South Eastern and Central districts of the country. There appeared to be a correlation between the occurrence of  P. malariae  infection with  P. vivax  infection, with the former only occurring in districts that had substantial  P. vivax  circulation. The median age among 2–12 year olds for  P. vivax  infection was 5 years (Mean 5.13 years, interquartile range 3–7 years). The odds of being infected with  P. vivax  decreased by 7 % for each year increase in age (OR 0.93, 95 % CI 0.87–1.00,  p  = 0.056).    Conclusion  We have confirmed low parasite rate of asymptomatic  Plasmodium  species in Botswana, with the exception of  P.vivax  which was unexpectedly high. This has implication for the elimination campaign so a follow up study is warranted to inform decisions on new strategies that take this evidence into account in the elimination campaign.

Molecular evidence of high rates of asymptomatic P. vivax infection and very low P. falciparum malaria in Botswana