Limit of blank and limit of detection of Plasmodium falciparum thick blood smear microscopy in a routine setting in Central Africa

BACKGROUND: Proper malaria diagnosis depends on the detection of asexual forms of Plasmodium spp. in the blood. Thick blood smear microscopy is the accepted gold standard of malaria diagnosis and is widely implemented. Surprisingly, diagnostic performance of this method is not well investigated and many clinicians in African routine settings base treatment decisions independent of microscopy results. This leads to overtreatment and poor management of other febrile diseases. Implementation of quality control programmes is recommended, but requires sustained funding, external logistic support and constant training and supervision of the staff. This study describes an easily applicable method to assess the performance of thick blood smear microscopy by determining the limit of blank and limit of detection. These two values are representative of the diagnostic quality and allow the correct discrimination between positive and negative samples. METHODS: Standard-conform methodology was applied and adapted to determine the limit of blank and the limit of detection of two thick blood smear microscopy methods (WHO and Lambaréné method) in a research centre in Lambaréné, Gabon. Duplicates of negative and low parasitaemia thick blood smears were read by several microscopists. The mean and standard deviation of the results were used to calculate the limit of blank and subsequently the limit of detection. RESULTS: The limit of blank was 0 parasites/μL for both methods. The limit of detection was 62 and 88 parasites/μL for the Lambaréné and WHO method, respectively. CONCLUSION: With a simple, back-of-the-envelope calculation, the performance of two malaria microscopy methods can be measured. These results are specific for each diagnostic unit and cannot be generalized but implementation of a system to control microscopy performance can improve confidence in parasitological results and thereby strengthen malaria control.