The Diagnostic Value of a Loop-Mediated Isothermal Amplification (LAMP) Technique for the Detection of Adenovirus Keratitis

Document Type : Original Article (s)

Authors

1 Department of Biology, Islamic Azad University, Science and Research Branch, Tehran, Iran

2 Associate Professor, Department of Microbiology, Islamic Azad University, Shahr-e-Qods Branch, Tehran, Iran

Abstract

Background: Adenovirus keratitis is an important ocular infection. Quick and timely detection of adenovirus keratitis can help to prevent the spread of disease. Virus detection methods (serological and molecular) have limitations. We evaluated the diagnostic value of a loop-mediated isothermal amplification (lamp) technique for the detection of adenovirus keratitis.Methods: In this descriptive cross-sectional study, 86 suspected patients to keratitis referred to Farabi and Labbafinejad Hospitals (Tehran, Iran) were enrolled in 2012. DNA samples of patients were extracted using boiling and DNG-Plus. Then, specific primers were designed for LAMP technique. First, sensitivity tests were done and then, optimized for the samples. At the end, LAMP production was examined adding cyber-green.Findings: The LAMP test was optimized regard to extracted DNA from adenovirus. The sensitivity test suggested the sensitivity about one particle per virus and no result was obtained during specification test by any of tested DNA. The results of 28 samples (32%) with specific particle and different titers were positive.Conclusion: Using LAMP technique to diagnose adenovirus showed that isothermal proliferation technique by loop (LAMP) has more sensitivity and precision. In addition, this technique provides a faster and more trusting method for diagnosing infections such as keratitis as an emergency case to be diagnosed and cured.

Keywords

References

  1. Aoki K, Tagawa Y. A twenty-one year surveillance of adenoviral conjunctivitis in Sapporo, Japan. Int Ophthalmol Clin 2002; 42(1): 49-54.
  2. Weitgasser U, Haller EM, El-Shabrawi Y. Evaluation of polymerase chain reaction for the detection of adenoviruses in conjunctival swab specimens using degenerate primers in comparison with direct immunofluorescence. Ophthalmologica 2002; 216(5): 329-32.
  3. Jawetz E, Melnick J. Medical Microbiology. New York, NY: McGraw-Hill Medical; 1980.
  4. Van RG, Klepper L, Peperkamp E, Schaap GJ. Immune electron microscopy and a cultural test in the diagnosis of adenovirus ocular infection. Br J Ophthalmol 1982; 66(5): 317-9.
  5. Roberts AW, Whitenack DL, Carter GR. Recovery of adenoviruses and slow herpesviruses from horses having respiratory tract infection. Am J Vet Res 1974; 35(9): 1169-72.
  6. Mori Y, Notomi T. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J Infect Chemother 2009; 15(2): 62-9.
  7. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 2000; 28(12): E63.
  8. Parida MM, Santhosh SR, Dash PK, Tripathi NK, Lakshmi V, Mamidi N, et al. Rapid and real-time detection of Chikungunya virus by reverse transcription loop-mediated isothermal amplification assay. J Clin Microbiol 2007; 45(2): 351-7.
  9. Mori Y, Nagamine K, Tomita N, Notomi T. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 2001; 289(1): 150-4.
  10. Nagamine K, Hase T, Notomi T. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 2002; 16(3): 223-9.
  11. Mori Y, Hirano T, Notomi T. Sequence specific visual detection of LAMP reactions by addition of cationic polymers. BMC Biotechnol 2006; 6: 3.
  12. Sohrabi N, Tebianian M, Moeini H. Analyzing bacterial agents of keratoconjunctivitis in patients referred to ophthalmology ward of Feiz hospital in Isfahan. J Fasa Univ Med Sci 2011; 2(1): 37-42. [In Persian].
  13. Kinchington PR, Turse SE, Kowalski RP, Gordon YJ. Use of polymerase chain amplification reaction for the detection of adenoviruses in ocular swab specimens. Invest Ophthalmol Vis Sci 1994; 35(12): 4126-34.
  14. Yoshida A, Nagashima S, Ansai T, Tachibana M, Kato H, Watari H, et al. Loop-mediated isothermal amplification method for rapid detection of the periodontopathic bacteria Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. J Clin Microbiol 2005; 43(5): 2418-24.
  15. Aonuma H, Yoshimura A, Perera N, Shinzawa N, Bando H, Oshiro S, et al. Loop-mediated isothermal amplification applied to filarial parasites detection in the mosquito vectors: Dirofilaria immitis as a study model. Parasit Vectors 2009; 2(1): 15.
  16. Reddy AK, Balne PK, Reddy RK, Mathai A, Kaur I. Development and evaluation of loop-mediated isothermal amplification assay for rapid and inexpensive detection of cytomegalovirus DNA in vitreous specimens from suspected cases of viral retinitis. J Clin Microbiol 2010; 48(6): 2050-2.
  17. Bahrami A, Shahhosseiny MH, Azadmanesh K, Noormohamadi Z, Moslemi E, Jadali F. Optimization of Loop-Mediated Isothermal Amplification (LAMP) method for detection of herpes simplex virus type 1 and 2. Sci J Kurdistan Univ Med Sci 2011, 16(1): 56-63. [In Persian].
  18. Esfahani SH, Shahhosseiny MH, Yaghmai P, Parivar K, Moslemi E, Keyvani Amini H. Rapid and simple detection of Hepatitis C virus by reverse transcriptase-loop-mediated isothermal amplification method. African Journal of Microbiology 2010; 4(23): 2580-6.
  19. Kohan L, Shahhosseiny MH, Razavi MR, Parivar K, Moslemi E, Werngren J. Evaluation of loop mediated isothermal amplification for diagnosis of Mycobacterium tuberculosis complex in clinical samples. African Journal of Biothechnology 2011; 10(26): 5096-101.
  20. Moradi A, Karami A, Hagh Nazari A, Ahmadi Z, Soroorizanjani R, Javadi SM. Comparison of the PCR and LAMP techniques in the diagnosis of Salmonella infection. J Zanjan Univ Med Sci 2009; 17(67): 66-77. [In Persian].
  21. Poschl B, Waneesorn J, Thekisoe O, Chutipongvivate S, Karanis P. Comparative diagnosis of malaria infections by microscopy, nested PCR, and LAMP in northern Thailand. Am J Trop Med Hyg 2010; 83(1): 56-60.
Journal of Isfahan Medical School
Volume 31, Issue 258 - Serial Number 258
September and October 2013
Pages 1713-1722

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History

  • Receive Date: 29 April 2013
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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