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Molecular Identification of Oxyspirura petrowi Intermediate Hosts by Nested PCR Using Internal Transcribed Spacer 1 (ITS1)

Molecular Identification of Oxyspirura petrowi Intermediate Hosts by Nested PCR Using Internal... ABSTRACTRecently, the heteroxenous eyeworm, Oxyspirura petrowi, has gained attention due to its prevalence in the declining game bird, Northern bobwhite (Colinus virginianus), but the intermediate hosts of many nematodes remain unknown. However, identifying the intermediate host of O. petrowi with traditional techniques would be difficult and time-consuming, especially considering there are more than 80 potential orthopteran hosts just in Texas. To screen a large number of samples quickly and effectively, primers for nested PCR (nPCR) were developed using the internal transcribed spacer 1 (ITS1) region. Then the nPCR was used to identify which of the 35 species collected from the Order Orthoptera were potential intermediate hosts of O. petrowi. With this technique, 18 potential intermediate hosts were identified. Later, we collected live specimens of species that tested positive to confirm the presence of larvae, but larvae were not found in the live specimens, nor in the extra tissue of the species that had tested positive for O. petrowi DNA. Despite this, this study demonstrated that nPCR is more sensitive than traditional techniques and can be a valuable tool in determining the intermediate hosts of parasites. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Parasitology Allen Press

Molecular Identification of Oxyspirura petrowi Intermediate Hosts by Nested PCR Using Internal Transcribed Spacer 1 (ITS1)

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The Journal of Parasitology , Volume 106 (1): 7 – Feb 28, 2020
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Publisher
Allen Press
Copyright
© American Society of Parasitologists 2020
ISSN
0022-3395
eISSN
1937-2345
DOI
10.1645/19-135
Publisher site
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Abstract

ABSTRACTRecently, the heteroxenous eyeworm, Oxyspirura petrowi, has gained attention due to its prevalence in the declining game bird, Northern bobwhite (Colinus virginianus), but the intermediate hosts of many nematodes remain unknown. However, identifying the intermediate host of O. petrowi with traditional techniques would be difficult and time-consuming, especially considering there are more than 80 potential orthopteran hosts just in Texas. To screen a large number of samples quickly and effectively, primers for nested PCR (nPCR) were developed using the internal transcribed spacer 1 (ITS1) region. Then the nPCR was used to identify which of the 35 species collected from the Order Orthoptera were potential intermediate hosts of O. petrowi. With this technique, 18 potential intermediate hosts were identified. Later, we collected live specimens of species that tested positive to confirm the presence of larvae, but larvae were not found in the live specimens, nor in the extra tissue of the species that had tested positive for O. petrowi DNA. Despite this, this study demonstrated that nPCR is more sensitive than traditional techniques and can be a valuable tool in determining the intermediate hosts of parasites.

Journal

The Journal of Parasitology – Allen Press

Published: Feb 28, 2020

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