Whatman Paper (FTA Cards) for Storing and Transferring Leishmania DNA for PCR Examination

Abstract

Background: Diagnosis of cutaneous leishmaniasis (CL) is often made based on clinical manifesta­tion. Correct diagnosis and identification of the parasite are crucial for choosing the effective treat­ment and for epidemiological studies. On the other hand, determination of Leishmania species is nec­essary for designing appropriate control programs. Diagnosis by PCR is becoming a 'gold standard'. For PCR preparation, storage and shipments of specimens are necessary. In this study, Whatman filter paper (FTA Card) was used to store and transfer samples for Leishmania identification using PCR.

Methods: Among the patients who had CL lesion and referred to Parasitology Laboratory of Emam Reza Hospital, Mashhad, Iran, 44 consented cases with positive results in their direct smear were se­lected. An informed consent form and a questionnaire were completed and three different types of samples (direct smear, NNN culture, and spot on FTA card) were collected. DNA extraction and PCR were carried out on three different samples from each patient.

Results: PCR results using Whatman paper samples revealed a significant difference (P<0.0001) compared to the culture method but no significant difference was seen between PCR results using samples stored on Whatman paper and direct smears.

Conclusion: The use of FTA cards is simple, rapid, and cost-effective, and can be readily employed for large-scale population screening, especially for regions where the specimens are to be transported from distant places to the laboratory.