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bioRxiv · 10.1101/2020.04.09.035147

A smartphone microscope method for simultaneous detection of (oo)cyst of Cryptosporodium and Giardia

Shrestha, R.·Dulal, R.·Wagle, S.·Pokhrel, S.·Giri, B.·Neupane, B. B.
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Abstract

Gastrointestinal disorders caused by ingestion of (oo)cysts of Cryptosporodium and Giardia is one of the major health problems in developing countries. We developed a smartphone based microscopic assay method to screen (oo)cysts of Cryptosporodium and Giardia contamination in vegetable and water samples. We used sapphire ball lens as the major imaging element to modify a smartphone as a microscope. Imaging parameters such as field of view and magnification, and image contrast under different staining and illumination conditions were measured. The smartphone microscope method consisting of ball lens of 1 mm diameter, white LED as illumination source and Lugolss iodine staining provided magnification and contrast capable of distinguishing (oo)cysts of Crypstopsporodium and Giardia in the same sample. The analytical performance of the method was tested by spike recovery experiments. The spiking recovery experiments performed on cabbage, carrot, cucumber, radish, tomatoes, and water resulted 26.8{+/-}10.3, 40.1{+/-}8.5, 44.4{+/-}7.3, 47.6{+/-}11.3, 49.2 {+/-}10.9, and 30.2{+/-}7.9% recovery for Cryptosporodium, respectively and 10.2{+/-}4.0, 14.1{+/-}7.3, 24.2{+/-}12.1, 23.2{+/-}13.7, 17.1{+/-}13.9, and 37.6{+/-}2.4 % recovery for Giardia, respectively. These recovery results were found to be similar when compared with the commercial brightfield and fluorescence microscopes. We tested the smartphone microscope system for detecting (oo)cysts on 7 types of vegetable (n=196) and river water (n=18) samples. Forty two percent vegetable and thirty-nine percent water samples were found to be contaminated with Cryptosporodium oocyst. Similarly, thirty one percent vegetable and thirty three percent water samples were contaminated with Giardia cyst. This study showed that the developed method can be a cheaper alternative for simultaneous detection of (oo)cysts in vegetable and water samples.

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Shrestha, R., Dulal, R., Wagle, S., Pokhrel, S., Giri, B., Neupane, B. B.. 2020-04-10. A smartphone microscope method for simultaneous detection of (oo)cyst of Cryptosporodium and Giardia. https://doi.org/10.1101/2020.04.09.035147

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