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Background   Aedes aegypti  and  Ae. albopictus  are important vectors of human diseases such as dengue, chikungunya, and zika. In Sri Lanka, they have been responsible for transmitting dengue virus. One of the most important parameters influencing the likelihood of arbovirus transmission is the age structure of the mosquito population. However, mosquito age is difficult to measure with accuracy. This study aims to construct multivariate calibration models using the transcriptional abundance of three age-responsive genes:  Ae15848  (calcium-binding protein),  Ae8505  (structural component of cuticle), and  Ae4274  (fizzy cell cycle/cell division cycle 20).    Methods  The transcriptional age-grading technique was applied to determine the chronological age of  Ae. aegypti  and  Ae. albopictus  female mosquito populations from Sri Lanka using the age-responsive genes  Ae15848 ,  Ae8505 , and  Ae4274 . Furthermore,  Ae. aegypti  samples obtained from colonies reared at two temperatures (23 and 27 °C) were used to investigate the influence of temperature on this age-grading technique. Expression levels of these three genes were quantified using reverse transcription qualitative PCR (qRT-PCR), and results were normalized against the housekeeping gene  ribosomal gene S17  ( RpS17 ).    Results  The expression of  Ae15848  and  Ae8505  decreased with the age of mosquitoes and showed the most significant and consistent change while expression of  Ae4274  increased with age. The multivariate calibration models showed > 80% correlation between expression of these age-responsive genes and the age of female mosquitoes at both temperatures. At 27 °C the accuracy of age predictions using the models was 2.19 (± 1.66) days and 2.58 (± 2.06) days for  Ae. aegypti  and  Ae. albopictus  females, respectively. The accuracy of the model for  Ae. aegypti  at 23 °C was 3.42 (± 2.74) days.    Conclusions  An adult rearing temperature difference of 4 °C (23–27 °C) did not significantly affect the age predictions. The calibration models created during this study could be successfully used to estimate the age of wild  Ae. aegypti  and  Ae. albopictus  mosquitoes from Sri Lanka.    Graphical Abstract

Use of transcriptional age grading technique to determine the chronological age of Sri Lankan Aedes aegypti and Aedes albopictus females