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-
1
World Health Organization, 1996. Fact Sheet No. 94. Geneva: World Health Organization.
-
2
Stauffer W, Fischer PR, 2003. Diagnosis and treatment of malaria in children. Clin Infect Dis 37 :1340–1348.
-
3
Amexo M, Tolhurst R, Barnish G, Bates I, 2004. Malaria misdiagnosis: effects on the poor and vulnerable. Lancet 364 :1896–1898.
-
4
Coleman R, Maneechat N, Rachaphaew N, Kumpitak C, Miller RS, Soyseng V, Thimasarn K, Sattabongkot J, 2002. Comparison of field and expert laboratory microscopy for active surveillance for asymptomatic Plasmodium falciparum and Plasmodium vivax in western Thailand. Am J Trop Med Hyg 67 :141–144.
-
5
McKenzie FE, Sirichaisinthop J, Miller RS, Gasser RA Jr, Wongsrichanalai C, 2003. Dependence of malaria detection and species diagnosis by microscopy on parasite density. Am J Trop Med Hyg 69 :372–376.
-
6
Ohrt C, Purnomo, Sutamihardja MA, Tang D, Kain KC, 2002. Impact of microscopy error on estimates of protective efficacy in malaria-prevention trials. J Infect Dis 186 :540–546.
-
8
Snounou G, Viriyakosol S, Jarra W, Thaithong S, Brown KN, 1993. Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections. Mol Biochem Parasitol 58 :283–292.
-
9
Arai M, Kunisada K, Kim H, Miyake H, Mizukoshi C, Kakutani T, Yamane A, Nakagami S, Kawai S, Kawamoto F, Wataya Y, 1996. A colorimetric DNA diagnostic method for falciparum malaria and vivax malaria: a field trial in the Solomon Islands. Nucleosides Nucleotides 15 :719–731.
-
10
Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA, 1999. A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg 60 :687–692.
-
11
Schindler HC, Montenegro L, Montenegro R, Carvalho AB, Abath FG, Jaureguiberry G, 2001. Development and optimization of polymerase chain reaction-based malaria diagnostic methods and their comparison with quantitative buffy coat assay. Am J Trop Med Hyg 65 :355–361.
-
12
Laoboonchai A, Kawamoto F, Thanoosingha N, Kojima S, Scott Miller RR, Kain KC, Wongsrichanalai C, 2001. PCR-based ELISA technique for malaria diagnosis of specimens from Thailand. Trop Med Int Health 6 :458–462.
-
13
McNamara DT, Thomson JM, Kasehagen LJ, Zimmerman PA, 2004. Development of a multiplex PCR-ligase detection reaction assay for diagnosis of infection by the four parasite species causing malaria in humans. J Clin Microbiol 42 :2403–2410.
-
14
Whiley DM, LeCornec GM, Baddeley A, Savill J, Syrmis MW, Mackay IM, Siebert DJ, Burns D, Nissen M, Sloots TP, 2004. Detection and differentiation of Plasmodium species by polymerase chain reaction and colorimetric detection in blood samples of patients with suspected malaria. Diagn Microbiol Infect Dis 49 :25–29.
-
15
Mockenhaupt FP, Ulmen U, von Gaertner C, Bedu-Addo G, Bienzle U, 2002. Diagnosis of placental malaria. J Clin Microbiol 40 :306–308.
-
16
Nsobya SL, Parikh S, Kironde F, Lubega G, Kamya MR, Rosenthal PJ, Dorsey G, 2004. Molecular evaluation of the natural history of asymptomatic parasitemia in Ugandan children. J Infect Dis 189 :2220–2226.
-
17
Lee MA, Tan CH, Aw LT, Tang CS, Singh M, Lee SH, Chia HP, Yap EP, 2002. Real-time fluorescence-based PCR for detection of malaria parasites. J Clin Microbiol 40 :4343–4345.
-
18
Farcas GA, Zhong KJ, Mazzulli T, Kain KC, 2004. Evaluation of the RealArt Malaria LC real-time PCR assay for malaria diagnosis. J Clin Microbiol 42 :636–638.
-
19
Rougemont M, van Saanen M, Sahli R, Hinrikson HP, Bille J, Jaton K, 2004. Detection of four Plasmodium species in blood from humans by 18S rRNA gene subunit-based and species-specific real-time PCR assays. J Clin Microbiol 42 :5636–5643.
-
20
Smith T, Uhl J, Espy M, Sloan L, Vetter E, Jones M, Rosenblatt J, Cockerill FI, 2004. Development, implementation, and trend analysis of real-time PCR tests for the clinical microbiology laboratory. Clin Microbiol Newsletter 26 :145–153.
-
21
Coyne SR, Craw PD, Norwood DA, Ulrich MP, 2004. Comparative analysis of the Schleicher and Schuell IsoCode STIX DNA isolation device and the Qiagen QIAamp DNA Mini Kit. J Clin Microbiol 42 :4859–4862.
-
22
Zhong KJ, Salas CJ, Shafer R, Gubanov A, Gasser RA Jr, Magill AJ, Forney JR, Kain KC, 2001. Comparison of IsoCode STIX and FTA Gene Guard collection matrices as whole-blood storage and processing devices for diagnosis of malaria by PCR. J Clin Microbiol 39 :1195–1196.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 135 | 95 | 9 |
| Full Text Views | 393 | 10 | 1 |
| PDF Downloads | 99 | 13 | 1 |
EVALUATION OF A REAL-TIME POLYMERASE CHAIN REACTION ASSAY FOR THE DIAGNOSIS OF MALARIA IN PATIENTS FROM THAILAND
HEATHER SWAN
HEATHER SWAN
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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LYNNE SLOAN
LYNNE SLOAN
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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ANTHONY MUYOMBWE
ANTHONY MUYOMBWE
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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PORNTIP CHAVALITSHEWINKOON-PETMITR
PORNTIP CHAVALITSHEWINKOON-PETMITR
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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SRIVICHA KRUDSOOD
SRIVICHA KRUDSOOD
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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WATTANA LEOWATTANA
WATTANA LEOWATTANA
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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POLRAT WILAIRATANA
POLRAT WILAIRATANA
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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SORNCHAI LOOAREESUWAN
SORNCHAI LOOAREESUWAN
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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JON ROSENBLATT
JON ROSENBLATT
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota; Department of Protozoology and Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
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We compared the diagnosis of malaria in 297 patients from Thailand by a real-time polymerase chain reaction (PCR) assay using the LightCycler with conventional microscopy using Giemsa-stained thick and thin blood films. The PCR assay can be completed in one hour and has the potential to detect and identify four species of Plasmodium in a single reaction by use of melting temperature curve analysis (however, we did not detect Plasmodium ovale in this study). Blood was collected, stored, and transported on IsoCode STIX, which provide a stable matrix for the archiving and rapid simple extraction of DNA. A genus-specific primer set corresponding to the 18S ribosomal RNA was used to amplify the target sequence. Fluorescence resonance energy technology hybridization probes were designed for P. falciparum over a region containing basepair mismatches, which allowed differentiation of the other Plasmodium species. The PCR results correlated with the microscopic results in 282 (95%) of 297 patient specimens. Most of these were single-species infections caused by P. vivax (150) and P. falciparum (120), along with 5 P. malariae, 2 mixed infections (P. falciparum and P. vivax), and 5 negative specimens. No negative microscopy specimens were positive by PCR (100% specificity for detection of any Plasmodium). The 15 discrepant results could not be resolved, but given the subjective nature of microscopy and the analytical objectivity of the PCR, the PCR results may be correct. The ability of the PCR method to detect mixed infections or to detect P. ovale could not be determined in this study. Within the limitations of initial equipment costs, this real-time PCR assay is a rapid, accurate, and efficient method for the specific diagnosis of malaria. It may have application in clinical laboratories, as well as in epidemiologic studies and antimalarial efficacy trials.
Author Notes
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-
1
World Health Organization, 1996. Fact Sheet No. 94. Geneva: World Health Organization.
-
2
Stauffer W, Fischer PR, 2003. Diagnosis and treatment of malaria in children. Clin Infect Dis 37 :1340–1348.
-
3
Amexo M, Tolhurst R, Barnish G, Bates I, 2004. Malaria misdiagnosis: effects on the poor and vulnerable. Lancet 364 :1896–1898.
-
4
Coleman R, Maneechat N, Rachaphaew N, Kumpitak C, Miller RS, Soyseng V, Thimasarn K, Sattabongkot J, 2002. Comparison of field and expert laboratory microscopy for active surveillance for asymptomatic Plasmodium falciparum and Plasmodium vivax in western Thailand. Am J Trop Med Hyg 67 :141–144.
-
5
McKenzie FE, Sirichaisinthop J, Miller RS, Gasser RA Jr, Wongsrichanalai C, 2003. Dependence of malaria detection and species diagnosis by microscopy on parasite density. Am J Trop Med Hyg 69 :372–376.
-
6
Ohrt C, Purnomo, Sutamihardja MA, Tang D, Kain KC, 2002. Impact of microscopy error on estimates of protective efficacy in malaria-prevention trials. J Infect Dis 186 :540–546.
-
8
Snounou G, Viriyakosol S, Jarra W, Thaithong S, Brown KN, 1993. Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections. Mol Biochem Parasitol 58 :283–292.
-
9
Arai M, Kunisada K, Kim H, Miyake H, Mizukoshi C, Kakutani T, Yamane A, Nakagami S, Kawai S, Kawamoto F, Wataya Y, 1996. A colorimetric DNA diagnostic method for falciparum malaria and vivax malaria: a field trial in the Solomon Islands. Nucleosides Nucleotides 15 :719–731.
-
10
Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA, 1999. A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg 60 :687–692.
-
11
Schindler HC, Montenegro L, Montenegro R, Carvalho AB, Abath FG, Jaureguiberry G, 2001. Development and optimization of polymerase chain reaction-based malaria diagnostic methods and their comparison with quantitative buffy coat assay. Am J Trop Med Hyg 65 :355–361.
-
12
Laoboonchai A, Kawamoto F, Thanoosingha N, Kojima S, Scott Miller RR, Kain KC, Wongsrichanalai C, 2001. PCR-based ELISA technique for malaria diagnosis of specimens from Thailand. Trop Med Int Health 6 :458–462.
-
13
McNamara DT, Thomson JM, Kasehagen LJ, Zimmerman PA, 2004. Development of a multiplex PCR-ligase detection reaction assay for diagnosis of infection by the four parasite species causing malaria in humans. J Clin Microbiol 42 :2403–2410.
-
14
Whiley DM, LeCornec GM, Baddeley A, Savill J, Syrmis MW, Mackay IM, Siebert DJ, Burns D, Nissen M, Sloots TP, 2004. Detection and differentiation of Plasmodium species by polymerase chain reaction and colorimetric detection in blood samples of patients with suspected malaria. Diagn Microbiol Infect Dis 49 :25–29.
-
15
Mockenhaupt FP, Ulmen U, von Gaertner C, Bedu-Addo G, Bienzle U, 2002. Diagnosis of placental malaria. J Clin Microbiol 40 :306–308.
-
16
Nsobya SL, Parikh S, Kironde F, Lubega G, Kamya MR, Rosenthal PJ, Dorsey G, 2004. Molecular evaluation of the natural history of asymptomatic parasitemia in Ugandan children. J Infect Dis 189 :2220–2226.
-
17
Lee MA, Tan CH, Aw LT, Tang CS, Singh M, Lee SH, Chia HP, Yap EP, 2002. Real-time fluorescence-based PCR for detection of malaria parasites. J Clin Microbiol 40 :4343–4345.
-
18
Farcas GA, Zhong KJ, Mazzulli T, Kain KC, 2004. Evaluation of the RealArt Malaria LC real-time PCR assay for malaria diagnosis. J Clin Microbiol 42 :636–638.
-
19
Rougemont M, van Saanen M, Sahli R, Hinrikson HP, Bille J, Jaton K, 2004. Detection of four Plasmodium species in blood from humans by 18S rRNA gene subunit-based and species-specific real-time PCR assays. J Clin Microbiol 42 :5636–5643.
-
20
Smith T, Uhl J, Espy M, Sloan L, Vetter E, Jones M, Rosenblatt J, Cockerill FI, 2004. Development, implementation, and trend analysis of real-time PCR tests for the clinical microbiology laboratory. Clin Microbiol Newsletter 26 :145–153.
-
21
Coyne SR, Craw PD, Norwood DA, Ulrich MP, 2004. Comparative analysis of the Schleicher and Schuell IsoCode STIX DNA isolation device and the Qiagen QIAamp DNA Mini Kit. J Clin Microbiol 42 :4859–4862.
-
22
Zhong KJ, Salas CJ, Shafer R, Gubanov A, Gasser RA Jr, Magill AJ, Forney JR, Kain KC, 2001. Comparison of IsoCode STIX and FTA Gene Guard collection matrices as whole-blood storage and processing devices for diagnosis of malaria by PCR. J Clin Microbiol 39 :1195–1196.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 135 | 95 | 9 |
| Full Text Views | 393 | 10 | 1 |
| PDF Downloads | 99 | 13 | 1 |