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Background  Malaria remains one of the leading communicable diseases in Ethiopia. Early diagnosis combined with prompt treatment is one of the main strategies for malaria prevention and control. Despite its limitation, Giemsa microscopy is still considered to be the gold standard for malaria diagnosis. This study aimed to compare the performance of Giemsa microscopy with nested polymerase chain reaction (nPCR) for the diagnosis of malaria in north-west Ethiopia.    Methods  A cross sectional study was conducted in public health facilities in North Gondar, from March 2013 to April 2013. A total number of 297 subjects with suspected malaria were enrolled in the study. Finger-prick blood samples were collected and examined for  Plasmodium  parasites using Giemsa microscopy and standard nPCR.    Results  Among the study participants, 61.6% (183/297) patients tested positive for malaria by Giemsa microscopy of which, 72.1% (132/183) and 27.9% (51/183) were diagnosed as  Plasmodium falciparum  and  Plasmodium vivax , respectively. By nPCR, 73.1% (217/297) were malaria-positive. Among microscopy-negative samples, 13.1% (39/297) samples turned malaria-positive in nPCR. In nPCR, the rate of mixed  Plasmodium  infections was 4.7% (14/297) and 3.03% (9/297) were positive for  Plasmodium ovale . Using nPCR as reference the sensitivity, specificity, positive predictive and negative predictive values of Giemsa microscopy were 82.0%, 93.8%, 97.3% and 65.8%, respectively, with a good agreement (κ = 0.668) to nested PCR. The sensitivity and specificity of Giemsa microscopy in identifying   P. falciparium  infections were 74.0% and 87.4% and 63.2% and 96.5% for  P. vivax  infections, respectively.    Conclusion  Although Giemsa microscopy remains the gold standard for malaria diagnosis in resource-limited environments, its sensitivity and specificity as compared to nPCR is limited suggesting exploration of novel rapid and simplified molecular techniques for malaria-endemic countries. A high rate of misclassification and misidentification highlights the importance of adequate training for staff involved in malaria diagnosis.

Comparison of Giemsa microscopy with nested PCR for the diagnosis of malaria in North Gondar, north-west Ethiopia