Dengue virus detection using whole blood for reverse transcriptase PCR and virus isolation.

Area of Science:

• Medical Virology
• Infectious Diseases
• Public Health

Background:

• Dengue poses a significant global health risk, affecting billions in tropical and subtropical areas.
• Accurate dengue virus detection is vital for patient management, disease surveillance, and clinical trials.
• Current diagnostic methods often rely on serum, requiring extensive sample processing.

Purpose of the Study:

• To compare the diagnostic efficacy of using whole blood versus serum/plasma for dengue virus detection.
• To evaluate the impact of different blood collection tubes (EDTA vs. serum separator tubes) on RT-PCR results.
• To assess the utility of whole blood in enhancing virus detection via cell culture and mosquito inoculation.

Main Methods:

• Reverse transcriptase PCR (RT-PCR) was performed on blood samples collected in EDTA tubes (whole blood) and serum separator tubes (serum/plasma).
• A total of 108 individuals with suspected dengue virus infection were included in the study.
• Virus detection was also assessed using direct C6/36 cell culture and mosquito inoculation methods.

Main Results:

• RT-PCR analysis of RNA extracted from whole blood demonstrated a higher dengue virus detection rate compared to serum or plasma.
• The use of whole blood samples in EDTA tubes improved diagnostic sensitivity.
• While whole blood also enhanced detection via cell culture and mosquito inoculation, the improvement was less pronounced than with RT-PCR.

Conclusions:

• Whole blood, particularly when collected in EDTA tubes, offers superior sensitivity for dengue virus detection using RT-PCR.
• This finding suggests that whole blood is a more advantageous sample type for acute dengue diagnosis.
• The enhanced sensitivity of whole blood in RT-PCR can improve clinical management and surveillance efforts for dengue virus infections.