Colorimetric LAMP PCR kit design for rapid diagnosis of Bovine coronavirus, Bovine rotavirus group A, Escherichia coli K99+ and Cryptosporidium parvum in Neonatal calf diarrhea

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Authors

  • Mehmet Demirci
  • Akın YIĞIN
  • Seda Ekici
  • Hikmet Dinç

Keywords:

Bovine coronavirus, Bovine rotavirus group A, Cryptosporidium parvum, Escherichia coli K99 , LAMP PCR, Neonatal calf diarrhea

Abstract

Reducing calf deaths and diseases are important for the future and profitability of cattle herds. Early diagnosis and treatment of calf diarrhea can reduce mortality rates and eliminate the need for cattle imports. It is important to produce local molecular kits based on the colorimetric Loop-mediated isothermal amplification (LAMP) method and to use them in the field, instead of using immunocrotographic tests with low sensitivity. With this study, four important enteric pathogens responsible for calf diarrhea (Bovine coronavirus [BCoV; formerly Betacoronavirus 1], group A Bovine rotavirus [BRV], Escherichia coli K99+ and Cryptosporidium parvum) were isolated in farm and field conditions without requiring qualified personnel and complicated devices. It is aimed to design a routine molecular kit based on the colorimetric LAMP method that can be detected in tubes in the range of 15-30 minutes. In addition, it is aimed to reveal the test kit performances by comparing the results of these LAMP PCR kits, which we will design, with in-house PCR and immunochromatographic test kits. A total of 100 calves stool samples were included (these samples were confirmed as positive for BcoV (n:25), BRV group A (n:25), Escherichia coli K99+ (n:25) and Cryptosporidium parvum (n:25) by in-house PCR). Nucleic acid (DNA/RNA) isolations from Neonatal Calf stool samples were performed using the commercial isolation kit. Results from LAMP kit and rapid antigen kits were compared according to in-house PCR results. Results of this study, especially E. coli K99 diagnostic performance were found to be closer to each other, while performance differences in viral parameters such as BRV and BcoV were found to be significantly different. It was determined that the LAMP PCR method performed better than rapid antigen test. As a conclusion, It is very important for early diagnosis to use the colorimetric LAMP method, which can be detected in separate tubes in 15-30 minutes in farm and field conditions without requiring qualified personnel and complicated devices. We believe that, thanks to early and reliable diagnosis, the calf mortality rates will be greatly reduced, as the infected animals will be treated early.

References

Al M ve Balıkçı E. 2012. Neonatal ishalli buzağılarda rotavirus, coronavirus, E. coli K99 ve Cryptosporidium parvum'un hızlı test kitleri ile teşhisi ve enteropatojen ile maternal immünite ilişkisi. FÜ Sağ. Bil. Vet. Derg, 26(2): 73-78.

Akbaş O, Yılmaz S, Başalan M. (2017. Aralık 21-22). Buzağı kayıpları sempozyumu. Kırıkkale Üniversitesi, Kırıkkale. Erişim adresi https://kku.edu.tr/Anasayfa/Etkinlik/Index/4517

Batmaz H. 2015. Buzağı Yönetimi ve Sağlığı. Editör: Batmaz H, Sığırlarda Sürü Sağlığı ve Yönetimi, 1. Baskı, Alfa Aktüel Yayınları, Bursa, s: 120-153

Bellino C, Arnaudo F, Biolatti C, Borrelli A, Gianella P, Maurella C, and D'Angelo A. 2012. Development of a diagnostic diagram for rapid field assessment of acidosis severity in diarrheic calves. Journal of the American Veterinary Medical Association, 240(3): 312-316.

Blanchard PC. 2012. Diagnostics of dairy and beef cattle diarrhea. Veterinary Clinics: Food Animal Practice. 28(3): 443-464.

Bilal T.2007. Yenidoğanların İç Hastalıkları. İstanbul Üniversitesi Veteriner Fakültesi, İstanbul. p. 158-69

Carter LJ, Garner LV, Smoot JW, et al. 2020. Assay Techniques and Test Development for COVID-19 Diagnosis. ACS Cent Sci. 6(5):591‐605.

Cho YI, Yoon KJ, 2014. An overview of calf diarrhea - infectious etiology, diagnosis, and intervention. J Vet Sci, 15:1-17.

Cho YI, Kim WI, Liu S, Kinyon JM, Yoon KJ. 2010. Development of a panel of multiplex real-time polymerase chain reaction assays for simultaneous detection of major agents causing calf diarrhea in feces. J Vet Diagn Invest. Jul;22(4):509-17.

Fan Q, Xie Z, Xie L, Liu J, Pang Y, Deng X, Xie Z, Peng Y, Wang X. A reverse transcription loop-mediated isothermal amplification method for rapid detection of bovine viral diarrhea virus. J Virol Methods. 2012;186(1-2):43-8.

Fischer S, Bauerfeind R, Czerny C et al. 2016. Serum interleukin-6 as a prognostic marker in a neonatal calf diarrhea. Journal of Dairy Science. 99(8): 6563-6571.

Foster DM, Smith GW. 2009. Pathophysiology of diarrhea in calves. Veterinary Clinics of North America: Food Animal Practice. 25(1):13-36.

Gambino CM, Lo Sasso B, Colomba C, et al. 2020. Comparison of a rapid immunochromatographic test with a chemiluminescence immunoassay for detection of anti-SARS-CoV-2 IgM and IgG. Biochem Med (Zagreb). 30(3):030901.

Glover AD, Puschner B, Rossow HA, Lehenbauer TW, Champagne JD, Blanchard PC, Aly SS. 2013. A double-blind block randomized clinical trial on the effect of zinc as a treatment for diarrhea in neonatal Holstein calves under natural challenge conditions. Prev Vet Med. 1;112(3-4):338-347.

Islam J, Tanimizu M, Shimizu Y, Goto Y, Ohtani N, Sugiyama K, Nochi T. 2022. Development of a rational framework for the therapeutic efficacy of fecal microbiota transplantation for calf diarrhea treatment. Microbiome, 10(1):1-20.

Karslı MA, Evci Ş. 2018. Buzağı Kayıplarının Önlenmesinde İnek ve Buzağı Beslemesinin Önemi. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi, 58: 23-34.

Kashir J, Yaqinuddin A. 2020. Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19 [published online ahead of print, 2020 Apr 25]. Med Hypotheses. 141:109786.

Kuliğ CC and Coşkun A. 2019. Sivas ve ilçelerindeki neonatal ishalli buzağılarda E. coli, Cryptosporidium, Clostridium perfringens, Rotavirüs ve Coronavirüs Prevalansı. Turkish Veterinary Journal, 1(2):69-73.

Macit, O. 2020. Tekirdağ ili süt sığırcılığı işletmelerinde buzağı ölümleri (Master's thesis, Fen Bilimleri Enstitüsü.

Maier GU, Breitenbuecher J, Gomez JP, Samah F, Fausak E, Van Noord M. 2022. Vaccination for the prevention of neonatal calf diarrhea in cow-calf operations: a scoping review. Veterinary and animal science, 100238.

Marino FE, Proffitt E, Joseph E, Manoharan A. 2022. A rapid, specific, extraction-less, and cost-effective RT-LAMP test for the detection of SARS-CoV-2 in clinical specimens. PLoS One. 11;17(4):e0266703.

Mawly JA, Grinberg A, Prattley D et al. 2015. Risk factors for neonatal calf diarrhoea and enteropathogen shedding in New Zealand dairy farms. Veterinary Journal 203(2): 155-160

Meganck V, Hoflack G, Opsomer G. 2014. Advances in prevention and therapy of neonatal dairy calf diarrhoea: a systemical rewiev with emphasis on colostrum management and fluid therapy. Acta Veterinaria Scandinavica 25; 56-75

Mendonça FL, Carvalho JG, Silva RJ, Ferreira LC, Cerqueira DM, Rogge HI, Facury-Filho EJ. 2021. Use of a natural herbal-based feed additive containing isoquinoline alkaloids in newborn calves with cryptosporidiosis. Veterinary parasitology, 300:109615.

Ok M, Güler L, Turgut K et al. 2009. The studies on the aetiology of diarrhoea in neonatal calves and determination of virulence gene markers of Escherichia coli strains by multiplex PCR. Zoonoses and Public Health 56:94-101.

Ozçelik A, Balıkcı E. 2018. Elazığ Yöresinde İshalli Neonatal Kuzu ve Oğlakların Bazı İshal Etkenlerinin Hızlı Test Kitleri İle Teşhisi. Fırat Üniversitesi Sağlık Bilimleri Veteriner Dergisi, 32(1):39-43.

Roubík H, Lošťák M, Ketuama CT, et al. 2022. Current coronavirus crisis and past pandemics - What can happen in post-COVID-19 agriculture?. Sustain Prod Consum. 30:752-760.

Sezer S and Akgul G. 2022. Rapıd Etıologıcal Dıagnosıs Of Neonatal Calf Dıarrhea Wıth Immunochromatographıc Test Kıts In Esme Dıstrıct Of Usak. Assiut Veterinary Medical Journal, 68(173):10-15.

Topal, O. 2018. Buzağılarda neonatal dönem sağlığını değerlendirmede ilk onbeş günde önemli olan klinik bulguların belirlenmesi. (Doktora tezi, Uludağ Üniv, Sağlıkbilimleri Enstitüsü Dahili Bilimler Anabilim Dalı)

TUIK,2021 SAYI 37208,13 Ağustos 2021

Xie Z, Fan Q, Liu J, Pang Y, Deng X, Xie Z, Liji X, Khan MI. Reverse transcription loop-mediated isothermal amplification assay for rapid detection of Bovine Rotavirus. BMC Vet Res. 2012;8:133.

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Published

2022-08-27

How to Cite

Demirci, M., Yigin, akın, Ekici, S., & Dinç, H. (2022). Colorimetric LAMP PCR kit design for rapid diagnosis of Bovine coronavirus, Bovine rotavirus group A, Escherichia coli K99+ and Cryptosporidium parvum in Neonatal calf diarrhea. International Journal of Veterinary and Animal Research (IJVAR), 5(2), 73–79. Retrieved from https://ijvar.org/index.php/ijvar/article/view/549

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Research Articles