Skip to main content
NCBI home page
Wiley - PMC COVID-19 Collection logoLink to Wiley - PMC COVID-19 Collection
. 2005 May 18;76(3):406–413. doi: 10.1002/jmv.20371

A simple method for the detection of measles virus genome by loop‐mediated isothermal amplification (LAMP)

Motoko Fujino 1, Naoko Yoshida 1, Shinya Yamaguchi 2, Norimitsu Hosaka 3, Yoshinori Ota 3, Tsugunori Notomi 3, Tetsuo Nakayama 1,✉
PMCID: PMC7166939  PMID: 15902708

Abstract

Approximately 20,000–30,000 measles patients were reported in a surveillance of infectious diseases because of low vaccine coverage of 80% in Japan. Among them, some were thought to be secondary vaccine failure (SVF) with generally mild or non‐typical measles illness and sometimes became a source of further transmission. We have developed a new, sensitive, and rapid method to detect the measles virus genome by reverse transcription loop‐mediated isothermal amplification (RT‐LAMP). We examined 50 nasopharyngeal secretion (NPS) samples that were obtained during the 1999 outbreak and stored at −70°C and fresh NPS, lymphocytes and sera from 11 patients in 2003. Total RNA was extracted from the samples and subjected to reverse transcription‐polymerase chain reaction (RT‐PCR) and RT‐LAMP. We detected the genomic RNA corresponding to at least 0.01–0.04 TCID50, 30–100 copies in samples by RT‐LAMP within 60 min after extraction of RNA, and all four genotypes isolated in Japan were equally amplified. Specific DNA amplification was monitored spectrophotometrically by real time turbidimeter and the quantity of RNA was calculated. Measles virus genome was detected in 44 of 50 stored NPS by RT‐PCR and in 49 by RT‐LAMP. The vaccine strain was discriminated from wild strains after sequencing the LAMP products. RT‐LAMP is a useful rapid diagnostic method for the detection of measles virus without any special apparatus, showing higher sensitivity than RT‐PCR, and expected to be applied for hospital‐based infection control and for laboratory‐based measles surveillance. J. Med. Virol. 76:406–413, 2005. © 2005 Wiley‐Liss, Inc.

Keywords: measles virus, reverse transcription‐polymerase reaction (RT‐PCR), reverse transcription‐loop‐mediated isothermal amplification (RT‐LAMP), rapid diagnosis

REFERENCES

  1. Afzal MA, Osterhaus ADME, Cosby SL, Jin L, Beeler J, Takeuchi K, Kawashima H. 2003. Comparative evaluation of measles virus‐specific RT‐PCR methods through an international collaborative study. J Med Virol 70: 171–176. [DOI] [PubMed] [Google Scholar]
  2. Griffin DE, Bellini WJ. 2001. Measles virus In: Fields virology, 4th edn. Philadelphia: Lippincott Williams & Wilkins; pp 1401–1441. [Google Scholar]
  3. Helfand RF, Kim DK, Gary HE, Jr. , Edwards GL, Bisson GP, Papania MJ, Heath JL, Schaff DL, Bellini WJ, Redd SC, Anderson LJ. 1998. Nonclassic measles infections in an immune population exposed to measles during a college bus trip. J Med Virol 56: 337–341. [DOI] [PubMed] [Google Scholar]
  4. Ihira M, Yoshikawa T, Enomoto Y, Akimoto S, Ohashi M, Suga S, Nishimura N, Ozaki T, Nishiyama Y, Notomi T, Ohta Y, Asano Y. 2004. Rapid diagnosis of human herpesvirus 6 infection by a novel DNA amplification method, loop‐mediated isothermal amplification. J Clin Microbiol 42: 140–145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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: 2616–2622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kobune F, Sakata H, Sugiura A. 1990. Marmoset lymphoblastoid cells as a sensitive host for isolation of measles virus. J Virol 64: 700–705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kuboki N, Inoue N, Sakurai T, Di Cello F, Grab DJ, Suzuki H, Sugimoto C, Igarashi I. 2003. Loop‐mediated isothermal amplification for detection of African trypanosomes. J Clin Microbiol 41: 5517–5524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kumada A, Komase K, Nakayana T. 2004. Recombinant measles AIK‐C strain expressing current wild‐type hemagglutinin protein. Vaccine 22: 309–316. [DOI] [PubMed] [Google Scholar]
  9. Lievano FA, Papania MJ, Helfand RF. 2004. Lack of evidence of measles virus shedding in people with inapparent measles virus infections J Infect Dis 189: S165–S170. [DOI] [PubMed] [Google Scholar]
  10. Maruyama F, Kenzaka T, Yamaguchi N, Tani K, Nasu M. 2003. Detection of bacteria carrying the stx2 gene by in situ loop‐mediated isothermal amplification. Appl Environ Microbiol 69: 5023–5028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mori Y, Kitao M, Tomita N, Notomi T. 2004. Real‐time turbidimetry of LAMP reaction for quantifying template DNA. J Biochem Biophys Methods 59: 145–157. [DOI] [PubMed] [Google Scholar]
  12. Mossong J, Nokes DJ, Edmunds WJ, Cox MJ, Ratnam S, Muller CP. 1999. Modeling the impact of inapparent measles transmission in vaccinated populations with waning immunity. Am J Epidemiol 150: 1238–1249. [DOI] [PubMed] [Google Scholar]
  13. Nagamine K, Hase T, Notomi T. 2002. Accelerated reaction by loop‐mediated isothermal amplification using loop primers. Mol Cell Probes 16: 223–229. [DOI] [PubMed] [Google Scholar]
  14. Nakayama T, Mori T, Yamaguchi S, Sonoda S, Asamura S, Yamashita R, Takeuchi Y, Urano T. 1995. Detection of measles virus genome directly from clinical samples by reverse transcriptase‐polymerase chain reaction and genetic variability. Virus Res 35: 1–16. [DOI] [PubMed] [Google Scholar]
  15. Nakayama T, Zhou J, Fujino M. 2003. Current status of measles in Japan. J Infect Chemother 9: 1–7. [DOI] [PubMed] [Google Scholar]
  16. Notomi T, Okayama H, Masubuchi H, Yonekawa T, K. Watanabe K, Amino N, Hase T. 2000. Loop‐mediated isothermal amplification of DNA. Nucleic Acids Res 28: e63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Okafuji T, Yoshida N, Fujino M, Motegi Y, Ihara T, Ota Y, Notomi T, Nakayama T. 2005. Rapid diagnostic method for the detection of mumps virus genome by loop‐mediated isothermal amplification. J Clin Microbiol 43: 1625–1631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Okamoto S, Yoshikawa T, Ihira M, Suzuki K, Shimokata K, Nishiyama Y, Asano Y. 2004. Rapid detection of varicella‐zoster virus infection by a loop‐mediated isothermal amplification method. J Med Virol 74: 677–682. [DOI] [PubMed] [Google Scholar]
  19. Ozoemena LC, Minor PD, Afzahl MA. 2004. Comparative evaluation of measles virus specific TaqMan PCR and conventional PCR using synthetic and natural RNA templates. J Med Virol 73: 79–84. [DOI] [PubMed] [Google Scholar]
  20. Parida M, Posadas G, Inoue S, Hasebe F, Morita K. 2004. Real‐time reverse transcription loop‐mediated isothermal amplification for rapid detection of West Nile virus. J Clin Microbiol 42: 257–263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schalk JAC, Van den Elzen C, Ovelgonne H, Baas C, Jongen PMJM. 2004. Estimation of the number of infectious measles viruses in live virus vaccine using real‐time PCR. J Virol Methods 117: 179–187. [DOI] [PubMed] [Google Scholar]
  22. Stein CE, Birmingham M, Kurian M, Duclos P, Strebel P. 2003. The global burden of measles in the year 2000—a model that uses country‐specific indicators. J Infect Dis 187: S8–S14. [DOI] [PubMed] [Google Scholar]
  23. Takahashi M, Nakayama T, Kashiwagi Y, Takami T, Sonoda S, Yamanaka T, Ochiai H, Ihara T, Tajima T. 2000. A single genotype of measles virus is dominant whereas several genotypes of mumps virus are co‐circulating. J Med Virol 62: 278–285. [PubMed] [Google Scholar]
  24. Thai HTC, Le MQ, Vuong CD, Parida M, Minekawa H, Notomi T, Hasebe F, Morita K. 2004. Development and evaluation of a novel loop‐mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus. J Clin Microbiol 42: 1956–1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. WHO . 2001. Standardization of the nomenclature for describing the genetic characteristics of wild‐type measles viruses. Wkly Epidemiol Rec 76: 241–248. [Google Scholar]
  26. WHO . 2002. WHO‐UNICEF joint statement on strategies to reduce measles mortality worldwide. Wkly Epidemiol Rec 77: 224–228. [PubMed] [Google Scholar]
  27. Wild TF, Malvoisin E, Buckland R. 1991. Measles virus: Both haemagglutinin and the fusion glycoproteins are required for fusion. J Gen Virol 72: 439–442. [DOI] [PubMed] [Google Scholar]
  28. Yamaguchi S. 1997. Identification of three lineages of wild measles virus by nucleotide sequence analysis of N, P, M, F and L genes in Japan. J Med Virol l 52: 113–120. [DOI] [PubMed] [Google Scholar]
  29. Yoshikawa T, Ihira M, Akimoto S, Usui C, Miyake F, Suga S, Enomoto Y, Suzuki Y, Nishiyama Y, Asano A. 2004. Detection of human herpesvirus 7 DNA by loop‐mediated isothermal amplification. J Clin Microbiol 42: 1348–1352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zhou J, Fujino M, Nakayama T. 2003. H1 genotype of measles virus was detected in outbreaks in Japan after 2000. J Med Virol 70: 642–648. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Medical Virology are provided here courtesy of Wiley

RESOURCES