Tracking false-negative results in molecular diagnosis: proposal of a triplex-PCR based method for leishmaniasis diagnosis

Gonçalves-Albuquerque, Suênia da CunhaSilva, Rômulo Pessoa eMorais, Rayana Carla Silva deTrajano-Silva, Lays Adrianne MendonçaRégis-da-Silva, Carlos GustavoBrandão-Filho, Sinval PintoPalva-Cavalcanti, Milena de

Molecular biological methods have become increasingly relevant to the diagnosis and control of infectious diseases, such as leishmaniasis. Since various factors may affect the sensitivity of PCR assays, including DNA yield and purity, an optimal extraction method is pivotal. Losses of a parasite’s DNA during extraction may significantly impair its detection by PCR and lead to false-negative results. This study proposes a triplex PCR assay targeting the parasite’s DNA, an external control (pUC18) and an internal control (G3PD) for accurate diagnosis of leishmaniasis.(AU)

1. pUC18
2. Leishmaniasis
3. Diagnosis
4. Extraction control
5. Multiplex PCR
6. False-negative result