Reply To: Development of malaria diagnosis in Anopheles mosquitoes

[Sakawrut Poosri]

1. The step of mosquito dissection for finding oocysts and sporozoite
1.1 Mosquito anesthetized by freezing at the -20 c for 5 min
1.2 Then, mosquito anesthesia, it will be pulled it’s leg and the body for easily to handle on the glass slide
1.3 Mosquito put on the glass slide
1.4 Use of two needles, first needle for fixing of abdomen, second needle use for pulling abdomen segment seven out of segment six
1.5 Then, we get the ovary, midgut, and malpighia tubule. This midgut to find the oocysts that embed midgut to epithelium microscope
1.6 We found the embed of oocytes at the midgut epithelium
1.7 The complete that oocytes contain a lot of sporozoite
But in the saliva gland for finding sporozoite, the process of mosquito dissection have different in step four. The needle use the two needles but for the first needle use for fixing the thorax and second needle use for pulled the head part. Then, we get the saliva gland and has three lobes
2. Mosquito dissection
Advantage: Easy to carry and handle, more specificity and use for gold standard. No need more equipment
Disadvantage: Using more time in this technique and low sensitivity and need expert to read the result
Immunological
Advantage: Easy to use this method and rapidly to detection and more specificity and sensitivity
Disadvantage: Expensive and unable to quantify parasite density
Molecular technique
Advantage: High sensitivity and specificity of detection, distinguish different spp., no need more time in this technique
Disadvantage: expensive, need more equipment and many machines, require highly train person to use method and not suitable for field
3. Using the mosquito dissection because this method is gold standard and high specificity and easy to handle. This method, it dose not have any equipment (microscope and expert to read the result). However, if we need to confirm the result, the sample will be contained to the laboratory for nested PCR that have more specificity and high sensitivity