Conference Abstract | Volume 8, Abstract ELIC2025212 (Poster 037) | Published: 31 Jul 2025
Sa’adatu Aliyu Abubakar1, &, Zacchaeus Adeniran Adejuyigbe1, Munzali Shamsu Zubairu1, Item Inya Item1, Yahaya Sakwa1, Adama Abubakar Ahmed1, Adesola Semiu Adeleye1, Mustapha Lawal1, Eugene Samuel Bwede1, Jessica Ewere Onyema1, Muraina Abbas Ayandayo1, Ikulughan Iibinmo Love1, James Avong1, Olajumoke Babatunde1, Jide Idris1
1Nigeria Centre for Disease Control and Prevention, Abuja, Nigeria
&Corresponding author: Sa’adatu Aliyu Abubakar, Nigeria Centre for Disease Control and Prevention, Abuja, Nigeria, Email: Saadatu.abubakar@ncdc.gov.ng
Received: 25 May 2025, Accepted: 09 Jun 2025, Published: 31 Jul 2025
Domain: Infectious Disease Epidemiology
Keywords: Lassa fever virus (LF), Ribonuclease enzyme (RNase), Real time (RT), Polymerase chain reaction (PCR), Cycle threshold (CT)
©Sa’adatu Aliyu Abubakar et al. Journal of Interventional Epidemiology and Public Health (ISSN: 2664-2824). This is an Open Access article distributed under the terms of the Creative Commons Attribution International 4.0 License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Cite this article: Sa’adatu Aliyu Abubakar et al., The effects of temperature variation and its impact on Lassa virus molecular detection at the national reference laboratory. Journal of Interventional Epidemiology and Public Health. 2025;8(ConfProc5):00181. https://doi.org/10.37432/JIEPH-CONFPRO5-00181
Lassa fever, poses a significant public health concern in West Africa, requiring accurate and timely detection, mainly using RT-PCR for effective management and outbreak control. In Nigeria, network of thirteen laboratories, including the National Reference Laboratory (NRL), conducts this testing. Samples often transported from remote areas and pass through several local logistics before reaching designated testing centres, which risks sample integrity. A key problem is temperature fluctuation during sample transport can trigger RNase activity, degrade viral RNA and compromise RT-PCR results. This study aims to investigate the effect of temperature variation impact on Lassa virus RNA detection by simulating real-world conditions commonly encountered during sample transport in Nigeria.
Eight PCR-positive samples were aliquoted into four groups and exposed to different temperature conditions for a period of five days as follows: Group 1: –15°C to –20°C (Freezer), Group 2: 2–8°C (Refrigerator), Group 3: ~25°C (Room Temperature) and Group 4: 28°C–33°C (Outdoor Conditions), At the end of the five-day period, all samples from each group were tested using RT-PCR.
The results were compared with the initial PCR results obtained prior to storage. A progressive increase in cycle threshold (Ct) values for both the GPC and L-gene targets was observed from Group 1 through Group 4. Group 1 showed the lowest Ct values, indicating optimal RNA preservation. In contrast, Group 4, recorded the highest Ct values, suggesting significant RNA degradation. Notably, in the L-gene targets for Groups 3 and 4, two out of eight samples yielded false-negative results, with no detectable L-gene amplification within the 45 cycles of PCR.
These findings emphasize the need to strengthen the pre-analytical phase of Lassa fever diagnostics, especially in decentralized settings where infrastructure and logistics may be limited. Appropriate sample handling and temperature control during transportation is essential for accurate detection and public health responses. Thus, have broader implications for the molecular detection of other RNA viruses in Nigeria, highlighting the necessity for a robust system that preserves Sample integrity.
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