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

Abstract

Abstract.

Diagnosis of gastrointestinal parasites has traditionally relied on stool microscopy, which has low diagnostic sensitivity and specificity. We have developed a novel, rapid, high-throughput quantitative multi-parallel real-time polymerase chain reaction (qPCR) platform. Species-specific primers/probes were used for eight common gastrointestinal parasite pathogens: , , , , spp., , , and . Stool samples from 400 13-month-old children in rural Ecuador were analyzed and the qPCR was compared with a standard direct wet mount slide for stool microscopy, as were 125 8–14-year-old children before and after anthelmintic treatment. The qPCR showed higher detection rates for all parasites compared with direct microscopy, (7.0% versus 5.5%) and for (31.5% versus 5.8%). Using an enhanced DNA extraction method, we were able to detect DNA. These assays will be useful to refine treatment options for affected populations, ultimately leading to better health outcomes.

Loading

Article metrics loading...

/content/journals/10.4269/ajtmh.12-0726
2013-06-05
2017-09-21
Loading full text...

Full text loading...

/deliver/fulltext/14761645/88/6/1041.html?itemId=/content/journals/10.4269/ajtmh.12-0726&mimeType=html&fmt=ahah

References

  1. Haque R, Mondal D, Karim A, Molla IH, Rahim A, Faruque AS, Ahmad N, Kirkpatrick BD, Houpt E, Snider C, Petri WA, Jr, 2009. Prospective case-control study of the association between common enteric protozoal parasites and diarrhea in Bangladesh. Clin Infect Dis 48: 11911197.[Crossref]
  2. ten Hove RJ, van Esbroeck M, Vervoort T, van den Ende J, van Lieshout L, Verweij JJ, , 2009. Molecular diagnostics of intestinal parasites in returning travellers. Eur J Clin Microbiol Infect Dis 28: 10451053.[Crossref]
  3. 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]
  4. Pecson BM, Barrios JA, Johnson DR, Nelson KL, , 2006. A real-time PCR method for quantifying viable Ascaris eggs using the first internally transcribed spacer region of ribosomal DNA. Appl Environ Microbiol 72: 78647872.[Crossref]
  5. Wolk DM, Schneider SK, Wengenack NL, Sloan LM, Rosenblatt JE, , 2002. Real-time PCR method for detection of Encephalitozoon intestinalis from stool specimens. J Clin Microbiol 40: 39223928.[Crossref]
  6. 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]
  7. Haque R, Neville LM, Hahn P, Petri WA, Jr, 1995. Rapid diagnosis of Entamoeba infection by using Entamoeba and Entamoeba histolytica stool antigen detection kits. J Clin Microbiol 33: 25582561.
  8. Petri WA, Jr, 2000. Protozoan parasites that infect the gastrointestinal tract. Cur Opin Gastroenterol 16: 1823.[Crossref]
  9. Haque R, Kabir M, Noor Z, Rahman SM, Mondal D, Alam F, Rahman I, Al Mahmood A, Ahmed N, Petri WA, Jr, 2010. Diagnosis of amebic liver abscess and amebic colitis by detection of Entamoeba histolytica DNA in blood, urine, and saliva by a real-time PCR assay. J Clin Microbiol 48: 27982801.[Crossref]
  10. Cooper PJ, Chico ME, Guadalupe I, Sandoval CA, Mitre E, Platts-Mills TA, Barreto ML, Rodrigues LC, Strachan DP, Griffin GE, , 2011. Impact of early life exposures to geohelminth infections on the development of vaccine immunity, allergic sensitization, and allergic inflammatory diseases in children living in tropical Ecuador: the ECUAVIDA birth cohort study. BMC Infect Dis 11: 184.[Crossref]
  11. Larson D, Cooper PJ, Hubner MP, Reyes J, Vaca M, Chico M, Kong HH, Mitre E, , 2012. Helminth infection is associated with decreased basophil responsiveness in human beings. J Allergy Clin Immunol 130: 270272.[Crossref]
  12. da Silva AJ, Bornay-Llinares FJ, Moura IN, Slemenda SB, Tutttle JL, Pieniazek NJ, , 1999. Fast and reliable extraction of protozoan parasite DNA from fecal specimens. Mol Diagn 4: 5764.[Crossref]
  13. Carlson JR, Sullivan PS, Harry DJ, Stork MA, Thorton SA, DuPont HL, , 1988. Enzyme immunoassay for the detection of Giardia lamblia . Eur J Clin Microbiol Infect Dis 7: 538540.[Crossref]
  14. Hotez P, , 2011. Enlarging the “audacious goal”: elimination of the world's high prevalence neglected tropical diseases. Vaccine 29 (Suppl 4): D104D110.[Crossref]
  15. Tejman-Yarden N, Eckmann L, , 2011. New approaches to the treatment of giardiasis. Curr Opin Infect Dis 24: 451456.[Crossref]
  16. Abu Al-Soud W, Radstrom P, , 1998. Capacity of nine thermostable DNA polymerases to mediate DNA amplification in the presence of PCR-inhibiting samples. Appl Environ Microbiol 64: 37483753.
  17. Koonjul PK, Brandt WF, Farrant JM, Lindsey GG, , 1999. Inclusion of polyvinylpyrrolidone in the polymerase chain reaction reverses the inhibitory effects of polyphenolic contamination of RNA. Nucleic Acids Res 27: 915916.[Crossref]
  18. Fahle GA, Fischer SH, , 2000. Comparison of six commercial DNA extraction kits for recovery of cytomegalovirus DNA from spiked human specimens. J Clin Microbiol 38: 38603863.
  19. Hotez PJ, Brooker S, Bethony JM, Bottazzi ME, Loukas A, Xiao S, , 2004. Hookworm infection. N Engl J Med 351: 799807.[Crossref]
  20. Bercu TE, Petri WA, Behm JW, , 2007. Amebic colitis: new insights into pathogenesis and treatment. Curr Gastroenterol Rep 9: 429433.[Crossref]
  21. Ungar BL, Soave R, Fayer R, Nash TE, , 1986. Enzyme immunoassay detection of immunoglobulin M and G antibodies to Cryptosporidium in immunocompetent and immunocompromised persons. J Infect Dis 153: 570578.[Crossref]
  22. Hoxie NJ, Davis JP, Vergeront JM, Nashold RD, Blair KA, , 1997. Cryptosporidiosis-associated mortality following a massive waterborne outbreak in Milwaukee, Wisconsin. Am J Public Health 87: 20322035.[Crossref]
  23. Nash TE, Herrington DA, Losonsky GA, Levine MM, , 1987. Experimental human infections with Giardia lamblia . J Infect Dis 156: 974984.[Crossref]
  24. Hall A, Nahar Q, , 1993. Albendazole as a treatment for infections with Giardia duodenalis in children in Bangladesh. Trans R Soc Trop Med Hyg 87: 8486.[Crossref]
  25. Basuni M, Mohamed Z, Ahmad M, Zakaria NZ, Noordin R, , 2012. Detection of selected intestinal helminths and protozoa at Hospital Universiti Sains Malaysia using multiplex real-time PCR. Trop Biomed 29: 434442.
  26. Petri WA, Jr Miller M, Binder HJ, Levine MM, Dillingham R, Guerrant RL, , 2008. Enteric infections, diarrhea, and their impact on function and development. J Clin Invest 118: 12771290.[Crossref]
http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.12-0726
Loading
/content/journals/10.4269/ajtmh.12-0726
Loading

Data & Media loading...

  • Received : 04 Dec 2012
  • Accepted : 18 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