Author:
ProCite
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Reference Manager
BibTeX
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-
1.
Caccio SM, Thompson RC, McLauchlin J, Smith HV, 2005. Unravelling Cryptosporidium and Giardia epidemiology. Trends Parasitol 21: 430–437.
-
2.
Savioli L, Smith H, Thompson A, 2006. Giardia and Cryptosporidium join the ‘Neglected Diseases Initiative.’ Trends Parasitol 22: 203–208.
-
3.
Checkley W, Gilman RH, Epstein LD, Suarez M, Diaz JF, Cabrera L, Black RE, Sterling CR, 1997. Asymptomatic and symptomatic cryptosporidiosis: their acute effect on weight gain in Peruvian children. Am J Epidemiol 145: 156–163.
-
4.
Prado MS, Cairncross S, Strina A, Barreto ML, Oliveira-Assis AM, Rego S, 2005. Asymptomatic giardiasis and growth in young children; a longitudinal study in Salvador, Brazil. Parasitology 131: 51–56.
-
5.
Mondal D, Haque R, Sack RB, Kirkpatrick BD, Petri WA Jr, 2009. Attribution of malnutrition to cause-specific diarrheal illness: evidence from a prospective study of preschool children in Mirpur, Dhaka, Bangladesh. Am J Trop Med Hyg 80: 824–826.
-
6.
Ajjampur SS, Sarkar R, Sankaran P, Kannan A, Menon VK, Muliyil J, Ward H, Kang G, 2010. Symptomatic and asymptomatic Cryptosporidium infections in children in a semi-urban slum community in southern India. Am J Trop Med Hyg 83: 1110–1115.
-
7.
Veenemans J, Mank T, Ottenhof M, Baidjoe A, Mbugi EV, Demir AY, Wielders JP, Savelkoul HF, Verhoef H, 2011. Protection against diarrhea associated with Giardia intestinalis is lost with multi-nutrient supplementation: a study in Tanzanian children. PLoS Negl Trop Dis 5: e1158.
-
8.
Elwin K, Robinson G, Hadfield SJ, Fairclough HV, Iturriza-Gomara M, Chalmers RM, 2012. A comparison of two approaches to extracting Cryptosporidium DNA from human stools as measured by a real-time PCR assay. J Microbiol Methods 89: 38–40.
-
9.
Schuurman T, Lankamp P, van Belkum A, Kooistra-Smid M, van Zwet A, 2007. Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens. Clin Microbiol Infect 13: 1186–1191.
-
10.
Verweij JJ, Schinkel J, Laeijendecker D, van Rooyen MA, van Lieshout L, Polderman AM, 2003. Real-time PCR for the detection of Giardia lamblia. Mol Cell Probes 17: 223–225.
-
11.
Verweij JJ, Blange RA, Templeton K, Schinkel J, Brienen EA, van Rooyen 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. J Clin Microbiol 42: 1220–1223.
-
12.
Robertson LJ, Gjerde BK, 2004. Effects of the Norwegian winter environment on Giardia cysts and Cryptosporidium oocysts. Microb Ecol 47: 359–365.
-
13.
Nichols RA, Moore JE, Smith HV, 2006. A rapid method for extracting oocyst DNA from Cryptosporidium-positive human faeces for outbreak investigations. J Microbiol Methods 65: 512–524.
-
14.
Haque R, Roy S, Siddique A, Mondal U, Rahman SM, Mondal D, Houpt E, Petri WA Jr, 2007. Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. Am J Trop Med Hyg 76: 713–717.
-
15.
Adamska M, Leońska-Duniec A, Maciejewska A, Sawczuk M, Skotarczak B, 2011. PCR and real time PCR for the detection of Cryptosporidium parvum oocyst DNA. Folia Biol (Krakow) 59: 115–120.
-
16.
Minarovicova J, Kaclikova E, Krascsenicsova K, Siekel P, Kuchta T, 2009. A single-tube nested real-time polymerase chain reaction for sensitive contained detection of Cryptosporidium parvum. Lett Appl Microbiol 49: 568–572.
-
17.
Mangani C, Maleta K, Phuka J, Cheung YB, Thakwalakwa C, Dewey K, Manary M, Puumalainen T, Ashorn P, 2013. Effect of complementary feeding with lipid-based nutrient supplements and corn-soy blend on the incidence of stunting and linear growth among 6- to 18-month-old infants and children in rural Malawi. Matern Child Nutr 25. doi:10.1111/mcn.12068.
-
18.
Adamska M, Leonska-Duniec A, Sawczuk M, Maciejewska A, Skotarczak B, 2012. Recovery of Cryptosporidium from spiked water and stool samples measured by PCR and real time PCR. Vet Med (Praha) 57: 224–232.
-
19.
Bruijnesteijn van Coppenraet LE, Wallinga JA, Ruijs GJ, Bruins MJ, Verweij JJ, 2009. Parasitological diagnosis combining an internally controlled real-time PCR assay for the detection of four protozoa in stool samples with a testing algorithm for microscopy. Clin Microbiol Infect 15: 869–874.
-
20.
Mary C, Chapey E, Dutoit E, Guyot K, Hasseine L, Jeddi F, Menotti J, Paraud C, Pomares C, Rabodonirina M, Rieux A, Derouin F, ANOFEL Cryptosporidium National Network, 2013. Multicentric evaluation of a new real-time PCR assay for quantification of Cryptosporidium spp. and identification of Cryptosporidium parvum and Cryptosporidium hominis. J Clin Microbiol 51: 2556–2563.
-
21.
ten Hove R, Schuurman T, Kooistra M, Moller L, van Lieshout L, Verweij JJ, 2007. Detection of diarrhoea-causing protozoa in general practice patients in the Netherlands by multiplex real-time PCR. Clin Microbiol Infect 13: 1001–1007.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 32 | 32 | 10 |
| Full Text Views | 354 | 144 | 0 |
| PDF Downloads | 126 | 33 | 0 |
High-Throughput Multiplex Quantitative Polymerase Chain Reaction Method for Giardia lamblia and Cryptosporidium Species Detection in Stool Samples
Noora Nurminen
Noora Nurminen
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Rosa Juuti
Rosa Juuti
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Sami Oikarinen
Sami Oikarinen
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Yue-Mei Fan
Yue-Mei Fan
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Kirsi-Maarit Lehto
Kirsi-Maarit Lehto
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Charles Mangani
Charles Mangani
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Kenneth Maleta
Kenneth Maleta
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Per Ashorn
Per Ashorn
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Heikki Hyöty
Heikki Hyöty
Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
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Giardia lamblia and Cryptosporidium species belong to a complex group of pathogens that cause diseases hampering development and socioeconomic improvements in the developing countries. Both pathogens are recognized as significant causes of diarrhea and nutritional disorders. However, further studies are needed to clarify the role of parasitic infections, especially asymptomatic infections in malnutrition and stunting. We developed a high-throughput multiplex quantitative polymerase chain reaction (qPCR) method for G. lamblia and Cryptosporidium spp. detection in stool samples. The sensitivity and specificity of the method were ensured by analyzing confirmed positive samples acquired from diagnostics laboratories and participating in an external quality control round. Its capability to detect asymptomatic G. lamblia and Cryptosporidium spp. infections was confirmed by analyzing stool samples collected from 44 asymptomatic 6-month-old infants living in an endemic region in Malawi. Of these, five samples were found to be positive for G. lamblia and two for Cryptosporidium spp. In conclusion, the developed method is suitable for large-scale studies evaluating the occurrence of G. lamblia and Cryptosporidium spp. in endemic regions and for clinical diagnostics of these infections.
Author Notes
Financial support: This study was financially supported by Academy of Finland (grants 200720, 108873, 111685, and 109796 [Per Ashorn]), Foundation for Pediatric Research in Finland, Nutriset Inc. (Malaunay, France), Päivikki and Sakari Sohlberg Foundation (Heikki Hyöty), Tampere Tuberculosis foundation (Heikki Hyöty). In addition, part of the study was funded by the DIABIMMUNE project (European Seventh Framework Programme HEALTH-F2-202063).
Authors' addresses: Noora Nurminen, Sami Oikarinen, and Heikki Hyöty, Department of Virology, School of Medicine, University of Tampere, Tampere, Finland, E-mails: noora.nurminen@uta.fi, sami.oikarinen@uta.fi, and heikki.hyoty@uta.fi. Rosa Juuti, EPID Research Oy, Espoo, Finland, E-mail: rosa.mattila@epidresearch.com. Yue-Mei Fan, Kirsi-Maarit Lehto, and Per Ashorn, Department for International Health, School of Medicine, University of Tampere, Tampere, Finland, E-mails: yuemei.fan@uta.fi, kirsi-maarit.lehto@uta.fi, and per.ashorn@uta.fi. Charles Mangani, Department for International Health, School of Medicine, University of Tampere, Tampere, Finland, and College of Medicine, University of Malawi, School of Public Health and Family Medicine, Blantyre, Malawi, E-mail: cmangani@medcol.mw. Kenneth Maleta, School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi, E-mail: kmaleta@medcol.mw.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Caccio SM, Thompson RC, McLauchlin J, Smith HV, 2005. Unravelling Cryptosporidium and Giardia epidemiology. Trends Parasitol 21: 430–437.
-
2.
Savioli L, Smith H, Thompson A, 2006. Giardia and Cryptosporidium join the ‘Neglected Diseases Initiative.’ Trends Parasitol 22: 203–208.
-
3.
Checkley W, Gilman RH, Epstein LD, Suarez M, Diaz JF, Cabrera L, Black RE, Sterling CR, 1997. Asymptomatic and symptomatic cryptosporidiosis: their acute effect on weight gain in Peruvian children. Am J Epidemiol 145: 156–163.
-
4.
Prado MS, Cairncross S, Strina A, Barreto ML, Oliveira-Assis AM, Rego S, 2005. Asymptomatic giardiasis and growth in young children; a longitudinal study in Salvador, Brazil. Parasitology 131: 51–56.
-
5.
Mondal D, Haque R, Sack RB, Kirkpatrick BD, Petri WA Jr, 2009. Attribution of malnutrition to cause-specific diarrheal illness: evidence from a prospective study of preschool children in Mirpur, Dhaka, Bangladesh. Am J Trop Med Hyg 80: 824–826.
-
6.
Ajjampur SS, Sarkar R, Sankaran P, Kannan A, Menon VK, Muliyil J, Ward H, Kang G, 2010. Symptomatic and asymptomatic Cryptosporidium infections in children in a semi-urban slum community in southern India. Am J Trop Med Hyg 83: 1110–1115.
-
7.
Veenemans J, Mank T, Ottenhof M, Baidjoe A, Mbugi EV, Demir AY, Wielders JP, Savelkoul HF, Verhoef H, 2011. Protection against diarrhea associated with Giardia intestinalis is lost with multi-nutrient supplementation: a study in Tanzanian children. PLoS Negl Trop Dis 5: e1158.
-
8.
Elwin K, Robinson G, Hadfield SJ, Fairclough HV, Iturriza-Gomara M, Chalmers RM, 2012. A comparison of two approaches to extracting Cryptosporidium DNA from human stools as measured by a real-time PCR assay. J Microbiol Methods 89: 38–40.
-
9.
Schuurman T, Lankamp P, van Belkum A, Kooistra-Smid M, van Zwet A, 2007. Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens. Clin Microbiol Infect 13: 1186–1191.
-
10.
Verweij JJ, Schinkel J, Laeijendecker D, van Rooyen MA, van Lieshout L, Polderman AM, 2003. Real-time PCR for the detection of Giardia lamblia. Mol Cell Probes 17: 223–225.
-
11.
Verweij JJ, Blange RA, Templeton K, Schinkel J, Brienen EA, van Rooyen 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. J Clin Microbiol 42: 1220–1223.
-
12.
Robertson LJ, Gjerde BK, 2004. Effects of the Norwegian winter environment on Giardia cysts and Cryptosporidium oocysts. Microb Ecol 47: 359–365.
-
13.
Nichols RA, Moore JE, Smith HV, 2006. A rapid method for extracting oocyst DNA from Cryptosporidium-positive human faeces for outbreak investigations. J Microbiol Methods 65: 512–524.
-
14.
Haque R, Roy S, Siddique A, Mondal U, Rahman SM, Mondal D, Houpt E, Petri WA Jr, 2007. Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. Am J Trop Med Hyg 76: 713–717.
-
15.
Adamska M, Leońska-Duniec A, Maciejewska A, Sawczuk M, Skotarczak B, 2011. PCR and real time PCR for the detection of Cryptosporidium parvum oocyst DNA. Folia Biol (Krakow) 59: 115–120.
-
16.
Minarovicova J, Kaclikova E, Krascsenicsova K, Siekel P, Kuchta T, 2009. A single-tube nested real-time polymerase chain reaction for sensitive contained detection of Cryptosporidium parvum. Lett Appl Microbiol 49: 568–572.
-
17.
Mangani C, Maleta K, Phuka J, Cheung YB, Thakwalakwa C, Dewey K, Manary M, Puumalainen T, Ashorn P, 2013. Effect of complementary feeding with lipid-based nutrient supplements and corn-soy blend on the incidence of stunting and linear growth among 6- to 18-month-old infants and children in rural Malawi. Matern Child Nutr 25. doi:10.1111/mcn.12068.
-
18.
Adamska M, Leonska-Duniec A, Sawczuk M, Maciejewska A, Skotarczak B, 2012. Recovery of Cryptosporidium from spiked water and stool samples measured by PCR and real time PCR. Vet Med (Praha) 57: 224–232.
-
19.
Bruijnesteijn van Coppenraet LE, Wallinga JA, Ruijs GJ, Bruins MJ, Verweij JJ, 2009. Parasitological diagnosis combining an internally controlled real-time PCR assay for the detection of four protozoa in stool samples with a testing algorithm for microscopy. Clin Microbiol Infect 15: 869–874.
-
20.
Mary C, Chapey E, Dutoit E, Guyot K, Hasseine L, Jeddi F, Menotti J, Paraud C, Pomares C, Rabodonirina M, Rieux A, Derouin F, ANOFEL Cryptosporidium National Network, 2013. Multicentric evaluation of a new real-time PCR assay for quantification of Cryptosporidium spp. and identification of Cryptosporidium parvum and Cryptosporidium hominis. J Clin Microbiol 51: 2556–2563.
-
21.
ten Hove R, Schuurman T, Kooistra M, Moller L, van Lieshout L, Verweij JJ, 2007. Detection of diarrhoea-causing protozoa in general practice patients in the Netherlands by multiplex real-time PCR. Clin Microbiol Infect 13: 1001–1007.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 32 | 32 | 10 |
| Full Text Views | 354 | 144 | 0 |
| PDF Downloads | 126 | 33 | 0 |