Hi Rashad,

as mainly the more mature stages can be found circulating, a common way of detection is using the Pfs25 QT-NASBA. 

Andrea

Hi Andrea,

Many thanks for your your kindness. I am a little bit confused about choosing a protocol for detection of Plasmodium falciparum  gametocytes. As a quantitative approach, QT-NASBA is the preferable choice. Is Pfs25 better than Pfs16? I would appreciate your help if you can send me a detailed protocol for QT-NASBA for P. falciparum gametocytes.

With my best regards,

Rashad

Dear Andrea,

What about using reverse transcriptase-PCR? Practically, what are the advantages and disadvantages of RT-PCR and QT-NASBA for the detection of P. falciparum gametocytes? 

Best regards,

Rashad

Hello My dear Rashed

Nested-PCR as a sensitive and specific method for  malaria diagnosis which can detect low parasite densities.

Best regards.

My dear Rashed Abdul- Ghani

Nested-PCR as a sensitive and specific method for  malaria diagnosis which can detect low parasite densities.

Best regards.

Thank you  for your contribution, dear Habibollah. For the detection of gametocytes, we have to target RNA rather than DNA. Therefore, we have to adopt strategies other than nested PCR.

Best regards,

Rashad

Hi Rashad,

apologies about the delayed reply. Pfs25 is currently the best way to detect late stage in the peripheral blood circulating gametocytes (as earlier stages are sequestered for example in the bone marrow). Have a look at this open access publication by Pritsch and colleagues "Stability of gametocyte-specific Pfs25-mRNA in dried blood spots on filter paper subjected to different storage conditions" (Malaria Journal). A detailed protocol can be found in there. I hope this helps and good luck with your work!

Kind regards,

Andrea