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Current IssueEditorial BoardOnline First ArticlesArchive

Short Communication

volume 52 issue 3 (march 2018) : 401-404,   Doi: 10.18805/ijar.B-567

First report on optimization of loop-mediated isothermal amplification (LAMP) for the diagnosis of Babesia felis

M
Muhammad Awais Salim
R
Raheela Akhtar
M
Muhammad Lateef
M
Muhammad Imran Rashid
H
Haroon Akbar
W
Wasim Shehzad
F
Fareeha Akhtar
S
Saher Islam
S
Shahid Hussain Farooqi
U
Uzma Farid Durrani
1Department of Parasitology, University of Veterinary and Animal Sciences, Lahore, Pakistan
Cite article:- Salim Awais Muhammad, Akhtar Raheela, Lateef Muhammad, Rashid Imran Muhammad, Akbar Haroon, Shehzad Wasim, Akhtar Fareeha, Islam Saher, Farooqi Hussain Shahid, Durrani Farid Uzma (2017). First report on optimization of loop-mediated isothermal amplification (LAMP) for the diagnosis of Babesia felis. Indian Journal of Animal Research. 52(3): 401-404. doi: 10.18805/ijar.B-567.

ABSTRACT

The objective of present study was to optimize loop mediated isothermal amplification (LAMP) assay for the diagnosis of Babesia felis in cats. LAMP primers were designed recognizing four sections of 18SribosomalRNA (18S rRNA) gene of B. felis. The blood samples of cats microscopically positive for Babesia felis were further used to extract deoxyribo neuclic acid (DNA) and the reaction mixture of 25 µL was standardized at 63°C temperature for 1 hour. LAMP assay provided more positive samples than conventional polymerase chain reaction (PCR). The prevalence of B. felis was also determined in cats using this optimized LAMP assay and it was found that the prevalence was more in younger cats as compare to adults. The application of LAMP can be helpful in rapid, reliable and cost effective diagnosis of B. felis in field.

REFERENCES

  1. Guan, G., Chauvin, A., Luo, J., Inoue, N., Moreau, E., Liu, Z., Gao, J., Thekisoe, O.M., Ma, M., Liu, A., Dang, Z., Liu, J., Ren, Q., Jin, Y., Sugimoto, C. and Yin, H. (2008). The development and evaluation of a loop-mediated isothermal amplification (LAMP) method for detection of Babesia spp. infective to sheep and goats in China. Exp. Parasitol. 120: 39-44.
  2. He, L., Zhou, Y.Q., Oosthuizen, M.C. and Zhao, J.L. (2009). Loop-mediated isothermal amplification (LAMP) detection of Babesia orientalis in water buffalo (Bubalus babalis) in China. Vet. Parasitol. 165: 36-40.
  3. Ikadai, H., Tanaka, H., Shibahara, N., Matsuu, A., Uechi, M., Itoh, N., Oshiro, S., Kudo, N., Igarashi, I. and Oyamada, T. (2004). Molecular evidence of infections with Babesia gibsoni parasites in Japan and evaluation of the diagnostic potential of a loop-mediated isothermal amplification method. J. Clin. Microbiol. 42: 2465-2469.
  4. Iseki, H., Alhassan, A., Ohta, N., Thekisoe, O.M., Yokoyama, N., Inoue, N., Nambota, A., Yasuda, J. and Igarashi, I. (2007). Development of a multiplex loopmediated isothermal amplification (mLAMP) method for the simultaneous detection of bovine Babesia parasites. J. Microbiol. Methods. 71: 281-287.
  5. Liu, A., Guan, G., Du, P., Gou, H., Liu, Z., Liu, J., Ma, M., Yang, J., Li, Y., Niu, Q., Ren, Q., Bai, Q., Yin, H. and Luo, J. (2012). Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina. Parasitol. Inst. 61: 658-663.
  6. Maia, C., Ramos, C., Coimbra, M., Bastos, F., Martins, A., Pinto, P., Nunes, M., Vieira, M.L., Cardoso, L. and Campino, L. (2014). Bacterial and protozoal agents of feline vector-borne diseases in domestic and stray cats from southern Portugal. Parasit. Vectors. 7:115-122.
  7. Makimura, K., Murayama, S.Y. and Yamaguchi, H. (1994). Detection of a wide range of medically important fungi by the polymerase chain reaction. J Med Microbiology. 40: 358-364.
  8. Monis, P.T., Giglio, S. and Saint, C.P. (2005). Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and investigation of the effect of dye concentration on amplification and DNA melting curve analysis. Anal. Biochem. 340: 24-34.
  9. Mori, Y., Kanda, H. and Notomi, T. (2013). Loop-mediated isothermal amplification (LAMP): recent progress in research and development. J. Infect. Chemother. 19: 404-411.
  10. Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N. and Hase, T. (2000). Loop-mediated isothermal amplification of DNA. Nucleic. Acids. Res. 28: E63.
  11. Pan, L., Zhang, L., Fan, D., Zhang, X., Liu, H., Lu, Q. and Xu, Q. (2013). Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. PLoS. Negl, Trop. Dis. 7: e2231.
  12. Parida, M., Sannarangaiah, S., Dash, P.K., Rao, P.V. and Morita, K. (2008). Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Rev. Med. Virol. 18: 407-421.
  13. Reddy, A.K., Balne, P.K., Reddy, R.K., Mathai, A. and Kaur, I. (2010). Development and evaluation of loop-lediated isothermal amplification assay for rapid and inexpensive detection of Cytomegalovirus DNA in vitreous specimens from suspected cases of viral retinitis. J. Clin. Microbiol. 48: 2050–2052.
  14. Tabar, M.D., Altet, L., Francino, O., Sanchez, A., Ferrer and L., Roura, X. (2008). Vector-borne infections in cats: molecular study in Barcelona area (Spain). Vet. Parasitol. 151: 332-336. 
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Indian Journal of Animal Research
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    Short Communication

    volume 52 issue 3 (march 2018) : 401-404,   Doi: 10.18805/ijar.B-567

    First report on optimization of loop-mediated isothermal amplification (LAMP) for the diagnosis of Babesia felis

    M
    Muhammad Awais Salim
    R
    Raheela Akhtar
    M
    Muhammad Lateef
    M
    Muhammad Imran Rashid
    H
    Haroon Akbar
    W
    Wasim Shehzad
    F
    Fareeha Akhtar
    S
    Saher Islam
    S
    Shahid Hussain Farooqi
    U
    Uzma Farid Durrani
    1Department of Parasitology, University of Veterinary and Animal Sciences, Lahore, Pakistan
    Cite article:- Salim Awais Muhammad, Akhtar Raheela, Lateef Muhammad, Rashid Imran Muhammad, Akbar Haroon, Shehzad Wasim, Akhtar Fareeha, Islam Saher, Farooqi Hussain Shahid, Durrani Farid Uzma (2017). First report on optimization of loop-mediated isothermal amplification (LAMP) for the diagnosis of Babesia felis. Indian Journal of Animal Research. 52(3): 401-404. doi: 10.18805/ijar.B-567.

    ABSTRACT

    The objective of present study was to optimize loop mediated isothermal amplification (LAMP) assay for the diagnosis of Babesia felis in cats. LAMP primers were designed recognizing four sections of 18SribosomalRNA (18S rRNA) gene of B. felis. The blood samples of cats microscopically positive for Babesia felis were further used to extract deoxyribo neuclic acid (DNA) and the reaction mixture of 25 µL was standardized at 63°C temperature for 1 hour. LAMP assay provided more positive samples than conventional polymerase chain reaction (PCR). The prevalence of B. felis was also determined in cats using this optimized LAMP assay and it was found that the prevalence was more in younger cats as compare to adults. The application of LAMP can be helpful in rapid, reliable and cost effective diagnosis of B. felis in field.

    REFERENCES

    1. Guan, G., Chauvin, A., Luo, J., Inoue, N., Moreau, E., Liu, Z., Gao, J., Thekisoe, O.M., Ma, M., Liu, A., Dang, Z., Liu, J., Ren, Q., Jin, Y., Sugimoto, C. and Yin, H. (2008). The development and evaluation of a loop-mediated isothermal amplification (LAMP) method for detection of Babesia spp. infective to sheep and goats in China. Exp. Parasitol. 120: 39-44.
    2. He, L., Zhou, Y.Q., Oosthuizen, M.C. and Zhao, J.L. (2009). Loop-mediated isothermal amplification (LAMP) detection of Babesia orientalis in water buffalo (Bubalus babalis) in China. Vet. Parasitol. 165: 36-40.
    3. Ikadai, H., Tanaka, H., Shibahara, N., Matsuu, A., Uechi, M., Itoh, N., Oshiro, S., Kudo, N., Igarashi, I. and Oyamada, T. (2004). Molecular evidence of infections with Babesia gibsoni parasites in Japan and evaluation of the diagnostic potential of a loop-mediated isothermal amplification method. J. Clin. Microbiol. 42: 2465-2469.
    4. Iseki, H., Alhassan, A., Ohta, N., Thekisoe, O.M., Yokoyama, N., Inoue, N., Nambota, A., Yasuda, J. and Igarashi, I. (2007). Development of a multiplex loopmediated isothermal amplification (mLAMP) method for the simultaneous detection of bovine Babesia parasites. J. Microbiol. Methods. 71: 281-287.
    5. Liu, A., Guan, G., Du, P., Gou, H., Liu, Z., Liu, J., Ma, M., Yang, J., Li, Y., Niu, Q., Ren, Q., Bai, Q., Yin, H. and Luo, J. (2012). Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina. Parasitol. Inst. 61: 658-663.
    6. Maia, C., Ramos, C., Coimbra, M., Bastos, F., Martins, A., Pinto, P., Nunes, M., Vieira, M.L., Cardoso, L. and Campino, L. (2014). Bacterial and protozoal agents of feline vector-borne diseases in domestic and stray cats from southern Portugal. Parasit. Vectors. 7:115-122.
    7. Makimura, K., Murayama, S.Y. and Yamaguchi, H. (1994). Detection of a wide range of medically important fungi by the polymerase chain reaction. J Med Microbiology. 40: 358-364.
    8. Monis, P.T., Giglio, S. and Saint, C.P. (2005). Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and investigation of the effect of dye concentration on amplification and DNA melting curve analysis. Anal. Biochem. 340: 24-34.
    9. Mori, Y., Kanda, H. and Notomi, T. (2013). Loop-mediated isothermal amplification (LAMP): recent progress in research and development. J. Infect. Chemother. 19: 404-411.
    10. Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N. and Hase, T. (2000). Loop-mediated isothermal amplification of DNA. Nucleic. Acids. Res. 28: E63.
    11. Pan, L., Zhang, L., Fan, D., Zhang, X., Liu, H., Lu, Q. and Xu, Q. (2013). Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. PLoS. Negl, Trop. Dis. 7: e2231.
    12. Parida, M., Sannarangaiah, S., Dash, P.K., Rao, P.V. and Morita, K. (2008). Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Rev. Med. Virol. 18: 407-421.
    13. Reddy, A.K., Balne, P.K., Reddy, R.K., Mathai, A. and Kaur, I. (2010). Development and evaluation of loop-lediated isothermal amplification assay for rapid and inexpensive detection of Cytomegalovirus DNA in vitreous specimens from suspected cases of viral retinitis. J. Clin. Microbiol. 48: 2050–2052.
    14. Tabar, M.D., Altet, L., Francino, O., Sanchez, A., Ferrer and L., Roura, X. (2008). Vector-borne infections in cats: molecular study in Barcelona area (Spain). Vet. Parasitol. 151: 332-336. 
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