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-
1
Van Eys GJJM, Schoone GJ, Kroon NCM, Ebeling SB, 1992. Sequence analysis of small subunit ribosomal RNA genes and its use for detection and identification of Leishmania parasites. Mol Biochem Parasitol 51 :133–142.
-
2
Cruz I, Canavate C, Rubio JM, Morales MA, Chicarro C, Laguna F, Jimenez-Mejias M, Sirera G, 2002. A nested polymerase chain reaction (Ln-PCR) for diagnosis and monitoring Leishmania infantum in patients coinfected with human immunodeficiency virus. Trans R Soc Trop Med Hyg 96 (Suppl 1):S185–S189.
-
3
Lachaud L, Dereure J, Chabbert E, Reynes J, Mauboussin JM, Oziol E, Dedet JP, Bastien P, 2000. Optimized PCR using patient blood samples for diagnosis and follow-up of visceral leishmaniasis with special reference to AIDS patients. J Clin Microbiol 38 :236–240.
-
4
Antinori S, Calattini S, Longhi E, Bestetti G, Piolini R, Magni C, Orlando G, Gramiccia M, Acquaviva V, Foschi A, Corvasce S, Colomba C, Titone L, Parravicini C, Cascio A, Corbellino M, 2007. Clinical use of polymerase chain reaction performed on peripheral blood and bone marrow samples for the diagnosis and monitoring of visceral leishmaniasis in HIV-infected and HIV-uninfected patients: a single-center, 8-year experience in Italy and review of the literature. Clin Infect Dis 44 :1602–1610.
-
5
Bourgeois N, Lachaud L, Reynes J, Rouanet I, Mahamat A, Bastien P, 2008. Long-term monitoring of visceral leishmaniasis in patients with AIDS. J Acquir Immune Defic Syndr 48 :13–19.
-
6
Higgins GA, 2001. Real-time PCR for the detection of Cryptosporidium parvum.J Microbiol Methods 47 :323–327.
-
7
Rolao N, Cortes S, Rodrigues OR, Campino L, 2004. Quantification of Leishmania infantum parasites in tissue biopsies by real-time polymerase chain reaction and polymerase chain reaction-enzyme-linked immunosorbent assay. J Parasitol 90 :1150–1154.
-
8
Blair PL, Witney A, Haynes JD, Moch JK, Carucci DJ, Adams JH, 2002. Transcripts of developmentally regulated Plasmodium falciparum genes quantified by real-time RT-PCR. Nucleic Acids Res 30 :2224–2231.
-
9
Croan DG, Morrison DA, Ellis JT, 1997. Evolution of the genus Leishmania revealed by comparison of DNA and RNA polymerase gene sequences. Mol Biochem Parasitol 89 :149–159.
-
10
Bretagne S, Durand R, Olivi M, Garin JF, Sulahian A, Rivollet D, Vidaud M, Deniau M, 2001. Real-time PCR as a new tool for quantifying Leishmania infantum in liver in infected mice. Clin Diagn Lab Immunol 8 :828–831.
-
11
Nicolas L, Prina E, Lang T, Milon G, 2002. Real-time PCR for detection and quantification of Leishmania in mouse tissues. J Clin Microbiol 40 :1666–1669.
-
12
Wortmann G, Hochberg L, Huong HH, Sweeney C, Zapor M, Aronson N, Weina P, Ockenhouse CF, 2005. Rapid identification of Leishmania complexes by a real-time PCR assay. Am J Trop Med Hyg 73 :999–1004.
-
13
Wortmann GW, Romero LI, Paz HM, Ortega-Barria E, Bayard V, Hochberg LP, Ryan JR, 2004. Real-time polymerase chain reaction diagnosis of leishmaniasis in Panama from both fresh and frozen tissue. Trans R Soc Trop Med Hyg 98 :148–151.
-
14
de Monbrison F, Mihoubi I, Picot S, 2007. Real-time PCR assay for the identification of cutaneous Leishmania parasite species in Constantine region of Algeria. Acta Trop 102 :79–83.
-
15
Schulz A, Mellenthin K, Schonian G, Fleischer B, Drosten C, 2003. Detection, differentiation, and quantitation of pathogenic Leishmania organisms by a fluorescence resonance energy transfer-based real-time PCR assay. J Clin Microbiol 41 :1529–1535.
-
16
Bossolasco S, Gaiera G, Olchini D, Gulletta M, Martello L, Bestetti A, Bossi L, Germagnoli L, Lazzarin A, Uberti-Foppa C, Cinque P, 2003. Real-time PCR assay for clinical management of human immunodeficiency virus-infected patients with visceral leishmaniasis. J Clin Microbiol 41 :5080–5084.
-
17
Mary C, Faraut F, Lascombe L, Dumon H, 2004. Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. J Clin Microbiol 42 :5249–5255.
-
18
Mary C, Faraut F, Drogoul MP, Xeridat B, Schleinitz N, Cuisenier B, Dumon H, 2006. References value for Leishmania infantum parasitemia in different clinical presentations: quantitative polymerase chain reaction for therapeutic monitoring and patient follow-up. Am J Trop Med Hyg 75 :858–863.
-
19
Sambrook JE, Fritsch F, Maniatis T, 1989. Molecular Cloning: A Laboratory Manual. Second edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
-
20
Minodier P, Piarroux R, Gambarelli F, Joblet C, Dumon H, 1997. Rapid identification of causative species in patients with Old World leishmaniasis. J Clin Microbiol 35 :2551–2555.
-
21
Lachaud L, Marchergui-Hammami S, Chabbert E, Dereure J, Dedet JP, Bastien P, 2002. Comparison of six PCR methods using peripheral blood for detection of canine visceral leishmaniasis. J Clin Microbiol 40 :210–215.
-
22
Aviles H, Belli A, Armijos R, Monroy FP, Harris E, 1999. PCR detection and identification of Leishmania parasites in clinical specimens in Ecuador: a comparison with classical diagnostic methods. J Parasitol 85 :181–187.
-
23
Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL, 2006. Comparison of PCR assays for diagnosis of cutaneous leishmaniasis. J Clin Microbiol 44 :1435–1439.
-
24
Bastien P, Procop GW, Reischl U, 2008. Quantitative real-time PCR is not more sensitive than “conventional” PCR. J Clin Microbiol 46 :1897–1900.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 235 | 176 | 12 |
| Full Text Views | 410 | 3 | 0 |
| PDF Downloads | 279 | 4 | 0 |
Is Real-Time Polymerase Chain Reaction (PCR) More Useful Than a Conventional PCR for the Clinical Management of Leishmaniasis?
Spinello Antinori
Spinello Antinori
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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Sara Calattini
Sara Calattini
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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Roberta Piolini
Roberta Piolini
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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Erika Longhi
Erika Longhi
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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Giovanna Bestetti
Giovanna Bestetti
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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Antonio Cascio
Antonio Cascio
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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Carlo Parravicini
Carlo Parravicini
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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Mario Corbellino
Mario Corbellino
Dipartimento di Scienze Cliniche L Sacco, sezione di Malattie Infettive e Immunopatologia, Università di Milano, Milano; Dipartimento di Patologia Umana, Unità Operatioa di Medicina Tropicale e Parassitologia, Università di Messina
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It is currently unknown if the use of a real-time polymerase chain reaction (PCR) adds value to the diagnosis and follow-up prognosis of patients affected by leishmaniasis. We performed a study using a real-time PCR directed against the α-polymerase gene and a semiquantitative PCR that target the SSU ribosomal RNA (rRNA) gene as control for the diagnosis and quantification of parasites in patients with visceral (VL) and cutaneous (CL) leishmaniasis. Our single copy real-time PCR missed one diagnosis of VL compared with the conventional PCR, whereas both PCR methods were able to detect Leishmania parasites in CL. Under anti-leishmania treatment the kinetics of parasitemia were comparable with the two methods. The real-time PCR directed against α-polymerase of Leishmania despite being able to make a more accurate quantification of parasites does not add to the decision-making management compared with a semiquantitative PCR, and it is comparatively expensive.
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-
1
Van Eys GJJM, Schoone GJ, Kroon NCM, Ebeling SB, 1992. Sequence analysis of small subunit ribosomal RNA genes and its use for detection and identification of Leishmania parasites. Mol Biochem Parasitol 51 :133–142.
-
2
Cruz I, Canavate C, Rubio JM, Morales MA, Chicarro C, Laguna F, Jimenez-Mejias M, Sirera G, 2002. A nested polymerase chain reaction (Ln-PCR) for diagnosis and monitoring Leishmania infantum in patients coinfected with human immunodeficiency virus. Trans R Soc Trop Med Hyg 96 (Suppl 1):S185–S189.
-
3
Lachaud L, Dereure J, Chabbert E, Reynes J, Mauboussin JM, Oziol E, Dedet JP, Bastien P, 2000. Optimized PCR using patient blood samples for diagnosis and follow-up of visceral leishmaniasis with special reference to AIDS patients. J Clin Microbiol 38 :236–240.
-
4
Antinori S, Calattini S, Longhi E, Bestetti G, Piolini R, Magni C, Orlando G, Gramiccia M, Acquaviva V, Foschi A, Corvasce S, Colomba C, Titone L, Parravicini C, Cascio A, Corbellino M, 2007. Clinical use of polymerase chain reaction performed on peripheral blood and bone marrow samples for the diagnosis and monitoring of visceral leishmaniasis in HIV-infected and HIV-uninfected patients: a single-center, 8-year experience in Italy and review of the literature. Clin Infect Dis 44 :1602–1610.
-
5
Bourgeois N, Lachaud L, Reynes J, Rouanet I, Mahamat A, Bastien P, 2008. Long-term monitoring of visceral leishmaniasis in patients with AIDS. J Acquir Immune Defic Syndr 48 :13–19.
-
6
Higgins GA, 2001. Real-time PCR for the detection of Cryptosporidium parvum.J Microbiol Methods 47 :323–327.
-
7
Rolao N, Cortes S, Rodrigues OR, Campino L, 2004. Quantification of Leishmania infantum parasites in tissue biopsies by real-time polymerase chain reaction and polymerase chain reaction-enzyme-linked immunosorbent assay. J Parasitol 90 :1150–1154.
-
8
Blair PL, Witney A, Haynes JD, Moch JK, Carucci DJ, Adams JH, 2002. Transcripts of developmentally regulated Plasmodium falciparum genes quantified by real-time RT-PCR. Nucleic Acids Res 30 :2224–2231.
-
9
Croan DG, Morrison DA, Ellis JT, 1997. Evolution of the genus Leishmania revealed by comparison of DNA and RNA polymerase gene sequences. Mol Biochem Parasitol 89 :149–159.
-
10
Bretagne S, Durand R, Olivi M, Garin JF, Sulahian A, Rivollet D, Vidaud M, Deniau M, 2001. Real-time PCR as a new tool for quantifying Leishmania infantum in liver in infected mice. Clin Diagn Lab Immunol 8 :828–831.
-
11
Nicolas L, Prina E, Lang T, Milon G, 2002. Real-time PCR for detection and quantification of Leishmania in mouse tissues. J Clin Microbiol 40 :1666–1669.
-
12
Wortmann G, Hochberg L, Huong HH, Sweeney C, Zapor M, Aronson N, Weina P, Ockenhouse CF, 2005. Rapid identification of Leishmania complexes by a real-time PCR assay. Am J Trop Med Hyg 73 :999–1004.
-
13
Wortmann GW, Romero LI, Paz HM, Ortega-Barria E, Bayard V, Hochberg LP, Ryan JR, 2004. Real-time polymerase chain reaction diagnosis of leishmaniasis in Panama from both fresh and frozen tissue. Trans R Soc Trop Med Hyg 98 :148–151.
-
14
de Monbrison F, Mihoubi I, Picot S, 2007. Real-time PCR assay for the identification of cutaneous Leishmania parasite species in Constantine region of Algeria. Acta Trop 102 :79–83.
-
15
Schulz A, Mellenthin K, Schonian G, Fleischer B, Drosten C, 2003. Detection, differentiation, and quantitation of pathogenic Leishmania organisms by a fluorescence resonance energy transfer-based real-time PCR assay. J Clin Microbiol 41 :1529–1535.
-
16
Bossolasco S, Gaiera G, Olchini D, Gulletta M, Martello L, Bestetti A, Bossi L, Germagnoli L, Lazzarin A, Uberti-Foppa C, Cinque P, 2003. Real-time PCR assay for clinical management of human immunodeficiency virus-infected patients with visceral leishmaniasis. J Clin Microbiol 41 :5080–5084.
-
17
Mary C, Faraut F, Lascombe L, Dumon H, 2004. Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. J Clin Microbiol 42 :5249–5255.
-
18
Mary C, Faraut F, Drogoul MP, Xeridat B, Schleinitz N, Cuisenier B, Dumon H, 2006. References value for Leishmania infantum parasitemia in different clinical presentations: quantitative polymerase chain reaction for therapeutic monitoring and patient follow-up. Am J Trop Med Hyg 75 :858–863.
-
19
Sambrook JE, Fritsch F, Maniatis T, 1989. Molecular Cloning: A Laboratory Manual. Second edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
-
20
Minodier P, Piarroux R, Gambarelli F, Joblet C, Dumon H, 1997. Rapid identification of causative species in patients with Old World leishmaniasis. J Clin Microbiol 35 :2551–2555.
-
21
Lachaud L, Marchergui-Hammami S, Chabbert E, Dereure J, Dedet JP, Bastien P, 2002. Comparison of six PCR methods using peripheral blood for detection of canine visceral leishmaniasis. J Clin Microbiol 40 :210–215.
-
22
Aviles H, Belli A, Armijos R, Monroy FP, Harris E, 1999. PCR detection and identification of Leishmania parasites in clinical specimens in Ecuador: a comparison with classical diagnostic methods. J Parasitol 85 :181–187.
-
23
Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL, 2006. Comparison of PCR assays for diagnosis of cutaneous leishmaniasis. J Clin Microbiol 44 :1435–1439.
-
24
Bastien P, Procop GW, Reischl U, 2008. Quantitative real-time PCR is not more sensitive than “conventional” PCR. J Clin Microbiol 46 :1897–1900.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 235 | 176 | 12 |
| Full Text Views | 410 | 3 | 0 |
| PDF Downloads | 279 | 4 | 0 |