Development and evaluation of a real-time PCR assay for quantification of Giardia and Cryptosporidium in sewage samples

Applied Microbiology and Biotechnology

Volumn 89, Issue 4, 2011, Pages 1203-1211

  Alonso, José L ^a   Amorós, Inmaculada ^a   Cañigral, Irene ^a  

^a UNIVERSITAT POLITÈCNICA DE VALÈNCIA   (Spain)

Author keywords

Cryptosporidium; Giardia; LightCycler; LNA probe; qPCR; Sewage; TaqMan

Indexed keywords

CRYPTOSPORIDIUM; GIARDIA; LIGHTCYCLER; LNA PROBE; QPCR; TAQMAN;

DNA; GENES; NUCLEIC ACIDS; PROBES; SEWAGE;

POLYMERASE CHAIN REACTION;

ASSAY; DIARRHEAL DISEASE; GENE EXPRESSION; GENETIC ANALYSIS; POLYMERASE CHAIN REACTION; PROTOZOAN; SENSITIVITY ANALYSIS; SEWAGE; SPECIES DIVERSITY; WATERBORNE DISEASE;

ARTICLE; CRYPTOSPORIDIUM; DNA EXTRACTION; GENE SEQUENCE; GIARDIA; NONHUMAN; NUCLEIC ACID PROBE; QUANTITATIVE ANALYSIS; REAL TIME POLYMERASE CHAIN REACTION; SAMPLING; SEWAGE;

CRYPTOSPORIDIUM; CYTOSKELETAL PROTEINS; GIARDIA; HUMANS; OLIGONUCLEOTIDE PROBES; PARASITOLOGY; POLYMERASE CHAIN REACTION; PROTOZOAN PROTEINS; SENSITIVITY AND SPECIFICITY; SEWAGE;

CRYPTOSPORIDIUM; GIARDIA; PROTOZOA;

EID: 79952534928     PISSN: 01757598     EISSN: None     Source Type: Journal    
DOI: 10.1007/s00253-010-2984-6     Document Type: Article

References (37)

1. Alonso JL, Amorós I, Cuesta G (2010) LNA probes in a real-time TaqMan PCR assay for genotyping of Giardia duodenalis in wastewaters. J Appl Microbiol 108:1594-1601
2. Bertrand I, Gantzer C, Chesnot T, Schwartzbrod J (2004) Improved specificity for Giardia lamblia cyst quantification in wastewater by development of a real-time PCR method. J Microbiol Methods 57:41-53 (Pubitemid 38299350)
3. Cacciò SM, De Giacomo M, Aulicino FA, Pozio E (2003) Giardia cysts in wastewater treatment plants in Italy. Appl Environ Microbiol 69:3393-3398 (Pubitemid 36693346)
4. Dupont HL, Chappell CL, Sterling CR, Okhuysen PC, Rose JB, Jakubowski W (1995) The infectivity of Cryptosporidium parvum in healthy volunteers. New Engl J Med 332:855-859
5. Fontaine M, Guillot E (2002) Development of a TaqMan quantitative PCR assay specific for Cryptosporidium parvum. FEMSMicrobiol Lett 214:13-17 (Pubitemid 35245516)
6. Fontaine M, Guillot E (2003) Study of 18S rRNA and rDNA stability by real-time RT-PCR in heat-inactivated Cryptosporidium parvum oocysts. FEMS Microbiol Lett 226:237-243 (Pubitemid 38444946)
7. Giovanni GD, LeChevallier MW (2005) Quantitative-PCR assessment of Cryptosporidium parvum cell culture infection. Appl Environ Microbiol 71:1495-1500 (Pubitemid 40396688)
8. Guy RA, Payment P, Krull UJ, Horgen PA (2003) Real-time PCR for quantification of Giardia and Cryptosporidium in environmental water samples and sewage. Appl Environ Microbiol 69:5178-5185 (Pubitemid 37150736)
9. Guy RA, Xiao C, Horgen PA (2004) Real-time PCR assay for detection and genotype differentiation of Giardia lamblia in stool specimens. J Clin Microbiol 42:3317-3320 (Pubitemid 38938102)
10. Heid CA, Stevens J, Livak KJ, Williams PM (1996) Real time quantitative PCR. Genome Res 6:986-994 (Pubitemid 26388091)
11. Higgins JA, Fayer R, Trout JM, Xiao L, Lal AA, Kerby S, Jenkins MC (2001) Real-time PCR for the detection of Cryptosporidium parvum. J Microbiol Methods 47:323-327
12. Ibekwe AM, Grieve CM (2003) Detection and quantification of Escherichia coli O157:H7 in environmental samples by real-time PCR. J Appl Microbiol 94:421-431 (Pubitemid 36315034)
13. Jiang J, Alderisio KA, Singh A, Xiao L (2005) Development of procedures for direct extraction of Cryptosporidium DNA from water concentrates for relief of PCR inhibitors. Appl Environ Microbiol 71:1135-1141 (Pubitemid 40396647)
14. Josefsen MH, Löfstrom C, Sommer HM, Hoorfar J (2009) Diagnostic PCR: comparative sensitivity of four probe chemistries. Mol Cell Probes 23:201-203
15. Karanis P, Kourenti KK, Smith HV (2007) Waterborne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt. J Water Health 5:1-38 (Pubitemid 46415958)
16. Keegan AR, Fanok S, Monis PT, Saint CP (2003) Cell culture- TaqMan PCR assay for evaluation of Cryptosporidium parvum disinfection. Appl Environ Microbiol 69:2505-2511 (Pubitemid 36560323)
17. Koshkin AA, Singh SK, Nielsen P, Rajwanshi VK, Kumar R, Meldgaard M, Olsen CE, Wengel J (1998) LNA (locked nucleic acids): synthesis of the adenine, cytosine, guanine, 5-methylcytosine, thymine and uracil bicyclonucleoside monomers, oligomerisation, and unprecedented nucleic acid recognition. Tetrahedron 54:3607-3630 (Pubitemid 28108183)
18. Kreader CA (1996) Relief of amplification inhibition in PCR with bovine serum albumin or T4 gene 32 protein. Appl Environ Microbiol 62:1102-1106
19. Kubota K, Ohashi A, Imachi H, Harada H (2006) Improved in situ hybridization efficiency with locked-nucleic-acid-incorporated DNA probes. Appl Environ Microbiol 72:5311-5317
20. Kumar R, Singh SK, Koshkin AA, Rajwanshi VK, Meldgaard M, Wengel J (1998) The first analogues of LNA (locked nucleic acids): phosphorohioate-LNA and 2OElig;thio-LNA. Bioorg Med Chem Lett 8:2219-2222 (Pubitemid 28561609)
21. Limor JR, Lal AA, Xiao L (2002) Detection and differentiation of Cryptosporidium parasites that are pathogenic for humans by real-time PCR. J Clin Microbiol 40:2335-2338 (Pubitemid 34747329)
22. Mackay IM, Mackay JK, Nissen MD, Sloots TP (2007) Real-time PCR: history and fluorogenic chemistries. In: Mackay IA (ed) Real-time PCR in microbiology from diagnosis to characterization. Caister Academic, Norfolk, pp 1-39
23. Malorny B, Hoorfar J (2005) Toward standardization of diagnostic PCR testing of fecal samples: lessons from the detections of salmonellae in pigs. J Clin Microbiol 43:3033-3037 (Pubitemid 40981298)
24. McDonald LM, Sargent K, Armson A, Thompson RC, Reynoldson JA (2002) The development of a real-time quantitative-PCR method for characterization of a Cryptosporidium parvum in vitro culturing system and assessment of drug efficacy. Mol Biochem Parasitol 121:279-282 (Pubitemid 34603413)
25. Nichols RA, Campbell BM, Smith HV (2003) Identification of Cryptosporidium spp. oocysts in United Kingdom noncarbonated natural mineral waters and drinking waters by using a modified nested PCR-restriction fragment length polymorphism assay. Appl Environ Microbiol 69:4183-4189 (Pubitemid 36870080)
26. Nitsche A (2007) Oligonucleotide design for in-house real-time PCR applications in microbiology. In: Mackay IA (ed) Real-time PCR in microbiology from diagnosis to characterization. Caister Academic, Norfolk, pp 41-69
27. Nolan T, Hands RE, Bustin SA (2006) Quantification of mRNA using real-time RT-PCR. Nat Protoc 1:1559-1582
28. Obika S, Nanbu D, Hari Y, Andoh J, Morio K, Doi T, Imanishi T (1997) Stability and structural features of the duplexes containing nucleoside analogues with a fixed N-type conformation, 2′-O,4′-C- methyleneribonucleosides. Tetrahedron Lett 39: 5401-5404 (Pubitemid 28295121)
29. Rendtorff RC (1954) The experimental transmission of human intestinal protozoan parasites. II. Giardia lamblia cysts given in capsules. Am J Hyg 59:209-220
30. Reynisson E, Josefsen MH, Krause M, Hoorfar J (2006) Evaluation of probe chemistries and platforms to improve the detection limit of real-time PCR. J Microbiol Methods 66:206-216 (Pubitemid 43903028)
31. Sanjay KS, Nielsen P, Koshkin AA, Wengel J (1998) LNA (locked nucleic acids): synthesis and high affinity nucleic acid recognition. Chem Commun 4:455-456 (Pubitemid 128675297)
32. Silahtaroglu AN, Tommerup N, Vissing H (2003) FISHing with locked nucleic acids (LNA): evaluation of different LNA/DNA mixmers. Mol Cell Probes 17:165-169 (Pubitemid 36999813)
33. Smith S, Vigilant L, Morin PA (2002) The effects of sequence length and oligonucleotide mismatches on 5′ exonuclease assay efficiency. Nucleic Acids Res 30:e111
34. Verweij JJ, Blangé RA, Templeton K, Schinkel J, Brienen EA, van Rooynen MA, van Lieshout L, Polderman AM (2004) Simultaneous detection of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum in fecal samples by using multiplex real-time PCR. Appl Environ Microbiol 42:1220-1223
35. Vester B, Wengel J (2004) LNA (locked nucleic acid): high-affinity targeting of complementary RNA and DNA. Biochemistry 43:13233-13241 (Pubitemid 39391124)
36. Wilhelm J, Pingoud A, Hahn M (2001) Comparison between Taq DNA polymerase and its Stoffel fragment for quantitative realtime PCR. Biotechniques 30:1052-1062 (Pubitemid 32429946)
37. Xiao L, Cama V (2006) Cryptosporidium and cryptosporidiosis. In: Ortega YR (ed) Foodborne parasites. Springer, New York, pp 57-108