Journal of Pharmacogenomics & Pharmacoproteomics

Diagnosis of vivax malaria: Precision and sensitivity

4^th International conference on Predictive, Preventive and Personalized Medicine & Molecular Diagnostics

September 22-23, 2016 Phoenix, USA

Maria de Fatima Ferreira-da-Cruz, Natalia K Almeida de Oliveira, Otacilio da Cruz, Leila Mendonca, Guilherme Werneck and Claudio Tadeu Daniel-Ribeiro

Instituto Oswaldo Cruz, Brazil
Universidade Estadual do Rio de Janeiro, Brazil

Posters & Accepted Abstracts: J Pharmacogenomics Pharmacoproteomics

Abstract:

The prompt diagnosis of plasmodial species for correct and effective patient treatment prevents transmission, reintroduction of malaria and the worsening of health condition of the patient. The PCR allows detecting and quantifying parasites below the detection threshold of microscopic examination. The PCR method for P. vivax detection standardized in our laboratory is effective for detecting infection but does not allow the quantification and a diagnosis as fast as the real time PCR format. Furthermore, its precision, comprising the repeatability and reproducibility parameters is unknown. Thus, our aim was to develop a real-time PCR assay with SYBR® Green and TaqMan® systems for the diagnosis of P. vivax malarial infection. Our experimental design included the construction of a standard curve with P. vivax DNA, cloned or not, to determine linearity; the setting of the lower detection limit and analytical sensitivity to measure sensitivity and intra assay variations (repeatability) and oscillations between assays, operators and equipment (reproducibility) to set precision. The performance of these parameters showed linearity of 4�?104 to 4 copies/μL with cloned DNA and 1�?104 to 1 parasite/μL with uncloned P. vivax DNA, quantification threshold of 1.77 and 0.94 and analytical sensitivity of 1.13 and 1.17 copies/μL for SYBR® Green and TaqMan® systems, respectively. When compared conventional PCR with real time one, the detection limit remained 0.00001 parasite/μL and the precision was maintained 100% with 0.1 parasite/μL in SYBR® Green and 1 parasite/μL with TaqMan® and conventional PCR. We conclude that the real-time PCR is the eligible methodology for the detection of P. vivax parasites, the TaqMan® system is the most indicated for quantitative assays and this methodology could be used to replace conventional PCR in reference laboratories for the diagnosis of vivax malaria.

Biography :

Email: mffcruz28@gmail.com