The Construction of Full-Length cDNA Library for Otodectes cynotis
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Otodectes cynotis (Hering, 1838) is the pathogen of otodectic mange distributed worldwide. The mite mainly infests carnivores and, sometimes, humans. However, due to the lack of cDNA library, research on its pathogenesis has been challenging.
To solve this problem, the present study first sampled O. cynotis mites from an infested cat from Xi’an, China, for RNA extraction. Then, the full-length cDNA library was constructed using the SMART technique. Finally, positive clones > 500 bp and Hsc70-5 gene fragment specifically amplified from the cDNA library were sequenced and analyzed to verify the library’s reliability.
Results showed that RNA extracted from 300 mites had good quality with a concentration of 149 ng/μl and OD260/OD280 of 1.99. The library satisfied the quality standard of a good library with a titer of 5.02 × 105 PFU/ml and a combination rate of 97.61%. In addition, clone 4 and Hsc70-5 showed 98.38% and 99.72% identity with Ef1-α and Hsc70-5 gene sequences of O. cynotis in GenBank, respectively.
The cDNA library of O. cynotis constructed here was successful and reliable, creating the basis for research on RNA sequencing and functional genes of O. cynotis.
KeywordsOtodectic mange cDNA library Quality evaluation Specific primer detection
This work was supported by the National Natural Science Foundation of China (Nos. 81471972; 81271856).
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 1.Ahn A.J., Oh D.S., Ahn K.S., Shin S.S. 2013. First feline case of otodectosis in the Republic of Korea and successful treatment with imidacloprid/moxidectin topical solution. The Korean Journal of Parasitology, 51, 125–128. https://doi.org/10.3347/kjp.2013.51.1.125 CrossRefPubMedPubMedCentralGoogle Scholar
- 2.Arther R.G., Davis W.L., Jacobsen J.A., Lewis V.A., Settje T.L. 2015. Clinical evaluation of the safety and efficacy of 10% imidacloprid + 2.5% moxidectin topical solution for the treatment of ear mite (Otodectes cynotis) infestations in dogs. Veterinary Parasitology, 210, 64–68. https://doi.org/10.1016/j.vetpar.2015.02.022 CrossRefPubMedGoogle Scholar
- 3.Beugnet F., Bourdeau P., Chalvet-Monfray K., Cozma V., Farkas R., Guillot J., Halos L., Joachim A., Losson B., Miró G., Otranto D., Renaud M., Rinaldi L. 2014. Parasites of domestic owned cats in Europe: co-infestations and risk factors. Parasites & Vectors, 7, 291. https://doi.org/10.1186/1756-3305-7-291 CrossRefGoogle Scholar
- 6.Davis W.L., Arther R.G., Settje T.S. 2007. Clinical evaluation of the efficacy and safety of topically applied imidacloprid plus moxidectin against ear mites (Otodectes cynotis) in client-owned cats. Parasitology Research, 101, S19–S24. https://doi.org/10.1007/s00436-007-0607-7 CrossRefGoogle Scholar
- 7.Fischer K., Holt D.C., Haruma P., Currie B.J., Walton S.F., Kem D.J. 2003. Generation and characterization of cDNA clones from Sarcoptes scabiei var. hominis for an expressed sequence tag library: identification of homologues of house dust mite allergens. The American Journal of Tropical Medicine and Hygiene, 68, 61–64.CrossRefPubMedGoogle Scholar
- 10.Kenyon F., Welsh M., Parkinson J., Whitton C., Blaxter M.L., Knox D.P. 2003. Expressed sequence tag survey of gene expression in the scab mite Psoroptes ovis–allergens, proteases and free-radical scavengers. Parasitology, 126, 451–460. https://doi.org/10.1017/s0031182003003044 CrossRefPubMedGoogle Scholar
- 13.Li Z.H., Li Z.Q., Qi Y.Y., Wang J. 2015. Diagnosis and therapy of otodectic mange in dogs. Contemporary Animal Husbandry, (6), 18–19Google Scholar
- 15.Liu G.Q., Liu P.H., Shen L.P., Tang J.Y., Hao J.P. 2000. Diagnose and treatment of otodectic mange in dogs. Chinese Journal of Veterinary Science and Technology, 30, 20.Google Scholar
- 18.Six R.H., Clemence R.G., Thomas C.A., Behan S., Boy M.G., Watson P., Benchaoui H.A., Clements P.J.M., Rowan T.G., Jernigan A.D. 2000. Efficacy and safety of selamectin against Sarcoptes scabiei on dogs and Otodectes cynotis on dogs and cats presented as veterinary patients. Veterinary Parasitology, 91, 291–309. https://doi.org/10.1016/s0304-4017(00)00300-9 CrossRefPubMedGoogle Scholar
- 20.Tomisawa H. 1999. A case of mite parasitized in the human external auditory canal. Jibi Inkoka Tokeibu Geka, 71, 276–278Google Scholar