1921
Volume 88, Issue 6
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Abstract.

The use of real-time polymerase chain reaction (PCR) for detection of in stool has recently been described. We compared five DNA extraction methods by using normal human stool spiked with and tested by using a real-time PCR. The PowerSoil kit was found to be the best technique in terms of sensitivity and ease of use. The PCR detected DNA extracted from one spiked larva diluted 10. The PowerSoil kit was then used to extract DNA from 160 human survey samples. All culture positive specimens with a high and moderate larval load were identified by real-time PCR, but only 15% of specimens with low larval load were positive. Specificity was greater than 99%. The combination of the PowerSoil kit and real-time PCR reliably detected high to moderate larval numbers of in stools but was less sensitive when the larval load was low.

Loading

Article metrics loading...

/content/journals/10.4269/ajtmh.12-0437
2013-06-05
2017-09-26
Loading full text...

Full text loading...

/deliver/fulltext/14761645/88/6/1048.html?itemId=/content/journals/10.4269/ajtmh.12-0437&mimeType=html&fmt=ahah

References

  1. Repetto SA, Duran PA, Lasala MB, Gonzalez-Cappa SM, , 2010. High rate of strongyloidosis infection, out of endemic area, in patients with eosinophilia and without risk of exogenous reinfections. Am J Trop Med Hyg 82: 10881093.[Crossref]
  2. Winsberg GR, Sonnenschein E, Dyer AR, Schnadig V, Bonilla E, , 1975. Prevalence of intestinal parasites in Latino residents of Chicago. Am J Epidemiol 102: 526532.[Crossref]
  3. Sato Y, Kobayashi J, Toma H, Shiroma Y, , 1995. Efficacy of stool examination for detection of Strongyloides infection. Am J Trop Med Hyg 53: 248250.
  4. Hirata T, Nakamura H, Kinjo N, Hokama A, Kinjo F, Yamane N, Fujita J, , 2007. Increased detection rate of Strongyloides stercoralis by repeated stool examinations using the agar plate culture method. Am J Trop Med Hyg 77: 683684.
  5. Ganesh S, Cruz RJ, Jr, 2011. Strongyloidiasis: a multifaceted disease. Gastroenterol Hepatol 7: 194196.
  6. van Doorn HR, Koelewijn R, Hofwegen H, Gilis H, Wetsteyn JC, Wismans PJ, Sarfati C, Vervoort T, van Gool T, , 2007. Use of enzyme-linked immunosorbent assay and dipstick assay for detection of Strongyloides stercoralis infection in humans. J Clin Microbiol 45: 438442.[Crossref]
  7. Rodrigues RM, de Oliveira MC, Sopelete MC, Silva DA, Campos DM, Taketomi EA, Costa-Cruz JM, , 2007. IgG1, IgG4, and IgE antibody responses in human strongyloidiasis by ELISA using Strongyloides ratti saline extract as heterologous antigen. Parasitol Res 101: 12091214.[Crossref]
  8. Olsen A, van Lieshout L, Marti H, Polderman T, Polman K, Steinmann P, Stothard R, Thybo S, Verweij JJ, Magnussen P, , 2009. Strongyloidiasis: the most neglected of the neglected tropical diseases? Trans R Soc Trop Med Hyg 103: 967972.[Crossref]
  9. Krolewiecki AJ, Ramanathan R, Fink V, McAuliffe I, Cajal SP, Won K, Juarez M, Di Paolo A, Tapia L, Acosta N, Lee R, Lammie P, Abraham D, Nutman TB, , 2010. Improved diagnosis of Strongyloides stercoralis using recombinant antigen-based serologies in a community-wide study in northern Argentina. Clin Vaccine Immunol 17: 16241630.[Crossref]
  10. Verweij JJ, Canales M, Polman K, Ziem J, Brienen EA, Polderman AM, van Lieshout L, , 2009. Molecular diagnosis of Strongyloides stercoralis in faecal samples using real-time PCR. Trans R Soc Trop Med Hyg 103: 342346.[Crossref]
  11. Kramme S, Nissen N, Soblik H, Erttmann K, Tannich E, Fleischer B, Panning M, Brattig N, , 2011. Novel real-time PCR for the universal detection of Strongyloides species. J Med Microbiol 60: 454458.[Crossref]
  12. Basuni M, Muhi J, Othman N, Verweij JJ, Ahmad M, Miswan N, Rahumatullah A, Aziz FA, Zainudin NS, Noordin R, , 2011. A pentaplex real-time polymerase chain reaction assay for detection of four species of soil-transmitted helminths. Am J Trop Med Hyg 84: 338343.[Crossref]
  13. Wilde J, Eiden J, Yolken R, , 1990. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol 28: 13001307.
  14. Valentine-Thon E, , 2002. Quality control in nucleic acid testing: where do we stand? J Clin Virol 25 (Suppl 3): S13S21.[Crossref]
  15. Boom R, Sol C, Weel J, Lettinga K, Gerrits Y, van Breda A, Wertheim-Van Dillen P, , 2000. Detection and quantitation of human cytomegalovirus DNA in faeces. J Virol Methods 84: 114.[Crossref]
  16. Beld M, Minnaar R, Weel J, Sol C, Damen M, van der Avoort H, Wertheim-van Dillen P, van Breda A, Boom R, , 2004. Highly sensitive assay for detection of enterovirus in clinical specimens by reverse transcription-PCR with an armored RNA internal control. J Clin Microbiol 42: 30593064.[Crossref]
  17. Fredricks DN, Smith C, Meier A, , 2005. Comparison of six DNA extraction methods for recovery of fungal DNA as assessed by quantitative PCR. J Clin Microbiol 43: 51225128.[Crossref]
  18. Garcia LS, , 2001. Diagnostic Medical Parasitology. Fourth edition. Washington, DC: American Society for Microbiology Press.
  19. Yu Z, Morrison M, , 2004. Improved extraction of PCR-quality community DNA from digesta and fecal samples. Biotechniques 36: 808812.
  20. Nylund L, Heilig HG, Salminen S, de Vos WM, Satokari R, , 2010. Semi-automated extraction of microbial DNA from stool for qPCR and phylogenetic microarray analysis. J Microbiol Methods 83: 231235.[Crossref]
  21. Marra NM, Chiuso-Minicucci F, Machado GC, Zorzella-Pezavento SF, Franca TG, Ishikawa LL, Amarante AF, Sartori A, Amarante MR, , 2010. Faecal examination and PCR to detect Strongyloides venezuelensis in experimentally infected Lewis rats. Mem Inst Oswaldo Cruz 105: 5761.[Crossref]
http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.12-0437
Loading
/content/journals/10.4269/ajtmh.12-0437
Loading

Data & Media loading...

  • Received : 19 Jul 2012
  • Accepted : 11 Feb 2013

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error