Even with the existence of a valid method to easily detect hotspots of malaria infection, including asymptomatic populations, a second limitation is our ability to diagnose infected individuals accurately. It is not uncommon for individuals to be infected with parasites that are sustained at microparasitaemic levels. The ability of current diagnostic tools to detect infections when parasite densities are low is poor and individuals negative for malaria by RDT or microscopy are still infectious to mosquitoes.[
21] The commonly used tools for malaria diagnosis for research purposes include microscopy, rapid diagnostic tests, measuring the immune response to plasmodium (serology), and various methods of DNA detection including polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), or nucleic acid sequence-based amplification (NASBA)[
22]. Although microscopy and RDT are the most commonly used methods of diagnosis, PCR can detect sub- patent infections and is frequently used for research purposes.[
23,
24]. However, the validity of PCR has not been adequately assessed for asymptomatic or microparasitaemic individuals: with a negative PCR result, what is the probability that a person is truly negative or is the negative due to insufficient sensitivity of the test? The analytical sensitivity of a nested PCR is often cited as being able to detect as few as two parasites per milliliter of whole blood[
25]. Although the lower limit of detection is impressive, the consistency of the results in microparasitaemic individuals has never been properly assessed, especially in field settings where sample collection and storage may not be optimum. Microparasitaemic individuals produce gametocytes and likely contribute to the infectious reservoir[
14]. However, individuals with infections below the sensitivity of PCR have received little attention and the probability that these individuals will produce and transmit gametocytes to mosquitoes, as well as their contribution to the infectious reservoir, has not been addressed.