Abstract:【Abstract】Objective To explore the detection limits of Plasmodium vivax from stained blood smears using nested PCR. Methods Serial dilutions (1:10, 1:100, 1:1000, 1:2000, 1:4000, 1:8000, 1:16000 and 1:32000) of a peripheral blood sample containing highdensity P. vivax were prepared using a blood sample drawn from a healthy person as diluent solution. Blood smears for each concentration were made with 5 μl sample for the thick blood film and 2ul for the thin blood filmStandard blood smears (at 9 degrees of dilution) were prepared using highdensity P. vivax from a peripheral blood sample. After microscopic examination, DNA was extracted from the thick blood film and thin blood film respectively and nested PCR were conducted in the same time. Results The detection threshold for P. vivax from the thick blood film was 1:16000 (08 parasite/uL). As for the thin blood film, the detection threshold was 1:100. Conclusion Nested PCR can be used as an alternative and useful tool for the detection of lowdensity P. vivax from stained blood smears, and it’s valuable for the molecular diagnosis of malaria.