Development and evaluation of the kit for detection of SARS-associated coronavirus RNA

Abstract


Aim. To develop a diagnostic kit for detection of SARS (severe acute respiratory syndrome)-related coronavirus RNA based on reverse transcription and polymerase chain reaction and to estimate its specificity and sensitivity.
Material and methods. 68 virus and bacterial cultures, 240 clinical samples from people without SARS symptoms and also 22 RNA samples from patients with SARS symptoms received during the epidemic in Beijing were used.
Results. The specificity of the kit was determined using animal coronaviruses and other bacterial and viral strains, causing acute respiratory and intestinal infections, and was shown to be 100%. The sensitivity of the kit in different clinical samples was 2.2' Iff genome equivalents of recombinant SARS RNA in I ml of the specimen. The kit was evaluated in the Institute of Microbiology and Epidemiology of Beijing (China) using SARS-cov viral suspension and clinical samples from patients with suspected SARS. It was shown that the kit was able to detect 10 TCID/50 ml of SARS-Cov virus. Testing of clinical samples from patients with suspected SARS showed that diagnostic sensitivity of the kit was 95%. Detection of the SARS-Cov RNA was more effective in feces compared to sputum 990 and 40%, respectively).
Conclusion. The kit "AmpliSens SARS" for qualitative detection of SARS-related coronavirus RNA by reverse transcription and polymerase chain reaction (PCR) in nasopharyngeal wash/aspirates, naso/ oropharyngeal swabs, plasma, and extract from feces has been developed in the Central Research Institute for Epidemiology of the RF Ministry of Health. The kit contains reagents for RNA isolation and purification, cDNA synthesis by reverse transcription ofRNA, for PCR and for electrophoretic analysis of amplified products. The kit also contains recombinant positive and internal control samples allowing to control efficiency of analysis and showed good analytical and diagnostic characteristics.

References

  1. Lee N., Hui D. S., Wu A. et al. A major outbreak of severe acute respiratory syndrome in Hong Kong. N. Engl. J. Med. 2003; 348: 1984-1994.
  2. Poutanen S. M., Low D. E., Henry B. et al. Identification of severe acute respiratory syndrome in Canada. N. Engl. J. Med. 2003; 348 (20): 1995-2005.
  3. Hsu L. Y., Lee С. С., Green J. A. et al. Severe acute respiratory syndrome in Singapore: clinical features of index patient and initial contacts. Emerg. Infect. Dis. 2003; 9 (6): 713-717.
  4. Murra M. A., Jones S. J. M., Astell С. R. et al. The genome sequence of the SARS-associated coronavirus. Science 2003; 300: 1399-1404.
  5. Uo L. K., Godeke G. J., Raamsman M. J. B. et al. Retargeting of coronavirus by substitution of the spike glycoprotein ectodomain: Crossing the host cell species barrier. J. Virol. 2000; 74 (3): 1393-1406.
  6. Virus taxonomy. Classification and nomenclature of viruses / van Regenmortel M. H. V. et al. New York: Academic Press; 2000.
  7. Rota P. A., Oberste M. S., Monroe S. S. et al. Characterization of a novel coronavirus associated with severe acute respiratory syndrome. Science 2003; 300: 1394-1399.
  8. Poland A. M., Vennema H., Foley J. E. et al. Two related strains of feline infectious peritonitis virus isolated from immunocompromised cats infected with a feline enteric coronavirus. J. Clin. Microbiol. 1996; 34 (2): 3180-3184.
  9. Peiris J. S. M., Chu С. М., Cheng V. С. С. Clinical progression and viral load in a community outbreak of coronavirus-associated SARS pneumonia: a prospective study. Lancet 2003; 36): 1767-1772.
  10. Worobey M., Holmes E. C. Evolutionary aspects of recombination in RNA viruses. J. Gen. Virol. 1999; 80: 2535-2543.
  11. Banner L. R., Lai M. M. Random nature of coronavirus RNA recombination in the absence of selection pressure. Virology 1991; 185 (1): 441-445.
  12. Boom R., Sol С. J. A., Salimans M. M. M. et al. Rapid and simple method for purification of nucleic acides. J. Clin. Microbiol. 1990; 28: 495-503.

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Copyright (c) 2004 Shipulin G.A., Yatsyshina S.В., Chulanov V.P., Markelov M.Y., Shipulina O.Y., Podkolzin А.Т., Astakhova T.S., Shishova A., Wuchun Cao -., Liu Wei -., Fan Baochang -.

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