1921
Volume 84, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Abstract.

There is a lack of diagnostic tests for leptospirosis in technology-restricted settings. We developed loop-mediated isothermal amplification (LAMP) specific for the 16S ribosomal RNA gene () of pathogenic and intermediate group species. The lower limit of detection was 10 genomic equivalents/reaction, and analytical specificity was high; we observed positive reactions for pathogenic/intermediate groups and negative reactions for non-pathogenic species and other bacterial species. We evaluated this assay in Thailand by using a case–control study of 133 patients with laboratory-proven leptospirosis and 133 patients with other febrile illnesses. Using admission blood, we found that the LAMP showed positive results in 58 of 133 cases (diagnostic sensitivity = 43.6, 95% confidence interval [CI] = 35.0–52.5) and in 22 of 133 controls (diagnostic specificity = 83.5, 95% CI = 76.0–89.3). Sensitivity was high for 39 patients who were culture positive for spp. (84.6, 95% CI = 69.5–94.1). The LAMP can provide an admission diagnosis in approximately half of patients with leptospirosis, but its clinical utility is reduced by a lower specificity.

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2017-09-26
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References

  1. Suttinont C, Losuwanaluk K, Niwatayakul K, Hoontrakul S, Intaranongpai W, Silpasakorn S, Suwancharoen D, Panlar P, Saisongkorh W, Rolain JM, Raoult D, Suputtamongkol Y, , 2006. Causes of acute, undifferentiated, febrile illness in rural Thailand: results of a prospective observational study. Ann Trop Med Parasitol 100: 363370.[Crossref]
  2. Merien F, Baranton G, Perolat P, , 1995. Comparison of polymerase chain reaction with microagglutination test and culture for diagnosis of leptospirosis. J Infect Dis 172: 281285.[Crossref]
  3. Smythe LD, Smith IL, Smith GA, Dohnt MF, Symonds ML, Barnett LJ, McKay DB, , 2002. A quantitative PCR (TaqMan) assay for pathogenic Leptospira spp. BMC Infect Dis 2: 13.[Crossref]
  4. Palaniappan RU, Chang YF, Chang CF, Pan MJ, Yang CW, Harpending P, McDonough SP, Dubovi E, Divers T, Qu J, Roe B, , 2005. Evaluation of lig-based conventional and real time PCR for the detection of pathogenic leptospires. Mol Cell Probes 19: 111117.[Crossref]
  5. Kositanont U, Rugsasuk S, Leelaporn A, Phulsuksombati D, Tantitanawat S, Naigowit P, , 2007. Detection and differentiation between pathogenic and saprophytic Leptospira spp. by multiplex polymerase chain reaction. Diagn Microbiol Infect Dis 57: 117122.[Crossref]
  6. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T, , 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28: E63.[Crossref]
  7. Kubo T, Agoh M, Mai LQ, Fukushima K, Nishimura H, Yamaguchi A, Hirano M, Yoshikawa A, Hasebe F, Kohno S, Morita K, , 2009. Development of a reverse transcription-loop-mediated isothermal amplification assay for detection of pandemic (H1N1), 2009, virus as a novel molecular method for diagnosis of pandemic influenza in resource-limited settings. J Clin Microbiol 48: 728735.[Crossref]
  8. Chantratita N, Meumann E, Thanwisai A, Limmathurotsakul D, Wuthiekanun V, Wannapasni S, Tumapa S, Day NP, Peacock SJ, , 2008. Loop-mediated isothermal amplification method targeting the TTS1 gene cluster for detection of Burkholderia pseudomallei and diagnosis of melioidosis. J Clin Microbiol 46: 568573.[Crossref]
  9. Nemoto J, Sugawara C, Akahane K, Hashimoto K, Kojima T, Ikedo M, Konuma H, Hara-Kudo Y, , 2009. Rapid and specific detection of the thermostable direct hemolysin gene in Vibrio parahaemolyticus by loop-mediated isothermal amplification. J Food Prot 72: 748754.[Crossref]
  10. Lin X, Chen Y, Lu Y, Yan J, Yan J, , 2009. Application of a loop-mediated isothermal amplification method for the detection of pathogenic Leptospira . Diagn Microbiol Infect Dis 63: 237242.[Crossref]
  11. Shang ES, Summers TA, Haake DA, , 1996. Molecular cloning and sequence analysis of the gene encoding LipL41, a surface-exposed lipoprotein of pathogenic Leptospira species. Infect Immun 64: 23222330.
  12. Morey RE, Galloway RL, Bragg SL, Steigerwalt AG, Mayer LW, Levett PN, , 2006. Species-specific identification of Leptospiraceae by 16S rRNA gene sequencing. J Clin Microbiol 44: 35103516.[Crossref]
  13. Slack AT, Kalambaheti T, Symonds ML, Dohnt MF, Galloway RL, Steigerwalt AG, Chaicumpa W, Bunyaraksyotin G, Craig S, Harrower BJ, Smythe LD, , 2008. Leptospira wolffii sp. nov., isolated from a human with suspected leptospirosis in Thailand. Int J Syst Evol Microbiol 58: 23052308.[Crossref]
  14. Levett PN, Morey RE, Galloway RL, Steigerwalt AG, , 2006. Leptospira broomii sp. nov., isolated from humans with leptospirosis. Int J Syst Evol Microbiol 56: 671673.[Crossref]
  15. Arzouni JP, Parola P, La Scola B, Postic D, Brouqui P, Raoult D, , 2002. Human infection caused by Leptospira fainei . Emerg Infect Dis 8: 865868.[Crossref]
  16. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG, , 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25: 48764882.[Crossref]
  17. Nicholas KB, Nicholas HB, Deerfield DWII, , 1997. GeneDoc: Analysis and Visualization of Genetic Variation. EMBNEW . NEWS 4: 14.
  18. Sonthayanon P, Chierakul W, Wuthiekanun V, Blacksell SD, Pimda K, Suputtamongkol Y, Pukrittayakamee S, White NJ, Day NP, Peacock SJ, , 2006. Rapid diagnosis of scrub typhus in rural Thailand using polymerase chain reaction. Am J Trop Med Hyg 75: 10991102.
  19. Wuthiekanun V, Chierakul W, Limmathurotsakul D, Smythe LD, Symonds ML, Dohnt MF, Slack AT, Limpaiboon R, Suputtamongkol Y, White NJ, Day NP, Peacock SJ, , 2007. Optimization of culture of Leptospira from humans with leptospirosis. J Clin Microbiol 45: 13631365.[Crossref]
  20. Stallman ND, , 1984. International Committee on Systematic Bacteriology Subcommittee on the Taxonomy of Leptospira: Minutes of the Meeting, 6 to 10 August 1982, Boston, Massachusetts. Int J Syst Bacteriol 34: 258259.[Crossref]
  21. Thaipadungpanit J, Wuthiekanun V, Chierakul W, Smythe LD, Petkanchanapong W, Limpaiboon R, Apiwatanaporn A, Slack AT, Suputtamongkol Y, White NJ, Feil EJ, Day NP, Peacock SJ, , 2007. A dominant clone of Leptospira interrogans associated with an outbreak of human leptospirosis in Thailand. PLoS Negl Trop Dis 1: e56.[Crossref]
  22. Han E-T, Watanabe R, Sattabongkot J, Khuntirat B, Sirichaisinthop J, Iriko H, Jin L, Takeo S, Tsuboi T, , 2007. Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol 45: 25212528.[Crossref]
  23. Iwamoto T, Sonobe T, Hayashi K, , 2003. Loop-mediated isothermal amplification for direct detection of Mycobacterium tuberculosis complex, M. avium, and M. intracellulare in sputum samples. J Clin Microbiol 41: 26162622.[Crossref]
  24. Dassanayake D, Wimalaratna H, Agampodi S, Liyanapathirana V, Piyarathna T, Goonapienuwala B, , 2009. Evaluation of surveillance case definition in the diagnosis of leptospirosis, using the microscopic agglutination test: a validation study. BMC Infect Dis 9: 48.[Crossref]
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  • Received : 24 Aug 2010
  • Accepted : 18 Nov 2010

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