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Published in: Malaria Journal 1/2012

Open Access 01-12-2012 | Research

Microscopy and molecular biology for the diagnosis and evaluation of malaria in a hospital in a rural area of Ethiopia

Authors: Maria A Santana-Morales, Raquel N Afonso-Lehmann, Maria A Quispe, Francisco Reyes, Pedro Berzosa, Agustin Benito, Basilio Valladares, Enrique Martinez-Carretero

Published in: Malaria Journal | Issue 1/2012

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Abstract

Background

Malaria is a leading public health problem in Ethiopia. Accurate diagnosis of Plasmodium infections is crucial for the reduction of malaria in tropical areas and for epidemiological studies. The role of light microscopy (LM) as gold standard has been questioned and, therefore, new molecular methods have been developed for the detection of Plasmodium species. The aim of the present work was to compare different malaria diagnostic methods in order to detect the most common species of Plasmodium and to broaden the knowledge of malaria prevalence in a hospital in a rural area in Ethiopia.

Methods

A cross-sectional survey of 471 individuals was carried out in a hospital in the rural area of Gambo (Ethiopia). Blood samples were prepared for microscopic observation and collected in filter paper for Seminested-Multiplex PCR (SnM-PCR) and real time PCR (qPCR) testing. The SnM-PCR was considered as the gold standard technique and compared with the rest. Thus, agreement between SnM-PCR and LM was determined by calculating Kappa Statistics and correlation between LM and qPCR quantification was calculated by pair-wise correlation co-efficient.

Results

Samples analysed by LM and SnM-PCR were positive for Plasmodium sp. 5.5% and 10.5%, respectively. Sensitivity was 52.2% by LM and 70% by qPCR. Correlation co-efficient between microscopy counts and qPCR densities for Plasmodium vivax was R2 = 0.586. Prevalence was estimated at 7% (95% CI: 4.7–9.3). Plasmodium vivax was the dominant species detected and the difference was statistically significant (χ2 = 5.121 p < 0.05). The highest prevalence of the parasite (10.9%) was observed in age groups under 15 years old.

Conclusion

Accurate malaria diagnostic methods have a great effect in the reduction of the number of malaria-infected individuals. SnM-PCR detection of malaria parasites may be a very useful complement to microscopic examination in order to obtain the real prevalence of each Plasmodium species. Although SnM-PCR shows that it is a good tool for the determination of Plasmodium species, today light microscopy remains the only viabletool for malaria diagnosis in developing countries. Therefore, re-inforcement in the training of microscopists is essential for making the correct diagnosis of malaria. Plasmodium vivax was the predominant species in Gambo, a meso-endemic area for this species.
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Literature
1.
go back to reference WHO: World Health Organization Expert Committee on Malaria. WHO Technical report series N° 892. 2000, Geneva, Switzerland, Geneva, Switzerland WHO: World Health Organization Expert Committee on Malaria. WHO Technical report series N° 892. 2000, Geneva, Switzerland, Geneva, Switzerland
2.
go back to reference Molineaux L: The epidemiology of human malaria as an explanation of its distribution, including some implications for its control. Malaria: Principles and Practice of Malariology. Volume 2. Edited by: Wernsdorfer WH, McGregor I. 1988, London: Churchill Livingston, 969-970. Molineaux L: The epidemiology of human malaria as an explanation of its distribution, including some implications for its control. Malaria: Principles and Practice of Malariology. Volume 2. Edited by: Wernsdorfer WH, McGregor I. 1988, London: Churchill Livingston, 969-970.
3.
go back to reference MacDonald G: The Epidemiology and Control of Malaria. 1957, London, New York, and Toronto: Oxford University Press MacDonald G: The Epidemiology and Control of Malaria. 1957, London, New York, and Toronto: Oxford University Press
4.
go back to reference Cambell CC: Challenges facing malaria therapy in Africa. J Infect Dis. 1991, 163: 1207-1211. 10.1093/infdis/163.6.1207.CrossRef Cambell CC: Challenges facing malaria therapy in Africa. J Infect Dis. 1991, 163: 1207-1211. 10.1093/infdis/163.6.1207.CrossRef
5.
go back to reference Ministry of Health: Malaria and other vector borne diseases prevention and control team. National Five Years Strategic Plan for Malaria Control in Ethiopia. 2001, Addis Ababa: MOH Ministry of Health: Malaria and other vector borne diseases prevention and control team. National Five Years Strategic Plan for Malaria Control in Ethiopia. 2001, Addis Ababa: MOH
6.
go back to reference Rosanas-Urgell A, Mueller D, Betuela I, Barnadas C, Iga J, Zimmerman PA, del Portillo HA, Silba P, Mueller I, Felger I: Comparison of diagnostic methods for the detection and quantification of the four sympatric Plasmodium species in field samples from Papua New Guinea. Malar J. 2010, 9: 361-10.1186/1475-2875-9-361.PubMedCentralCrossRefPubMed Rosanas-Urgell A, Mueller D, Betuela I, Barnadas C, Iga J, Zimmerman PA, del Portillo HA, Silba P, Mueller I, Felger I: Comparison of diagnostic methods for the detection and quantification of the four sympatric Plasmodium species in field samples from Papua New Guinea. Malar J. 2010, 9: 361-10.1186/1475-2875-9-361.PubMedCentralCrossRefPubMed
7.
go back to reference WHO: World malaria report 2008. 2008, Geneva, Switzerland: World Health OrganizationPress WHO: World malaria report 2008. 2008, Geneva, Switzerland: World Health OrganizationPress
8.
go back to reference Moody AH, Chiodini PL: Non-microscopic method for malaria diagnosis using OptiMAL IT, a second-generation dipstick for malaria pLDH antigen detection. Br J Biomed Sci. 2002, 59: 228-231.PubMed Moody AH, Chiodini PL: Non-microscopic method for malaria diagnosis using OptiMAL IT, a second-generation dipstick for malaria pLDH antigen detection. Br J Biomed Sci. 2002, 59: 228-231.PubMed
9.
go back to reference Coleman RE, Sattabongkot J, Promstaporm S, Maneechai N, Tippayachai B, Kengluecha A, Rachapaew N, Zollner G, Miller RS, Vaughan JA, Thimasarn K, Khuntirat B: Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plasmodium falciparum/vivax endemic area in Thailand. Malar J. 2006, 5: 121-10.1186/1475-2875-5-121.PubMedCentralCrossRefPubMed Coleman RE, Sattabongkot J, Promstaporm S, Maneechai N, Tippayachai B, Kengluecha A, Rachapaew N, Zollner G, Miller RS, Vaughan JA, Thimasarn K, Khuntirat B: Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plasmodium falciparum/vivax endemic area in Thailand. Malar J. 2006, 5: 121-10.1186/1475-2875-5-121.PubMedCentralCrossRefPubMed
10.
go back to reference Ochola LB, Vounatsou P, Smith T, Mabaso ML, Newton CR: The reliability of diagnostic techniques in the diagnosis and management of malaria in the absence of a gold standard. Lancet Infect Dis. 2006, 6: 582-588. 10.1016/S1473-3099(06)70579-5.CrossRefPubMed Ochola LB, Vounatsou P, Smith T, Mabaso ML, Newton CR: The reliability of diagnostic techniques in the diagnosis and management of malaria in the absence of a gold standard. Lancet Infect Dis. 2006, 6: 582-588. 10.1016/S1473-3099(06)70579-5.CrossRefPubMed
11.
go back to reference Valenstein PN: Evaluating diagnostic tests with imperfect standards. Am J Clin Path. 1990, 93: 252-258.PubMed Valenstein PN: Evaluating diagnostic tests with imperfect standards. Am J Clin Path. 1990, 93: 252-258.PubMed
12.
go back to reference Kain KC, Lanar DE: Determination of genetic variation within Plasmodium falciparum by using enzymatically amplified DNA from filter paper disks impregnated with whole blood. J ClinMicrobiol. 1991, 29: 1171-1174. Kain KC, Lanar DE: Determination of genetic variation within Plasmodium falciparum by using enzymatically amplified DNA from filter paper disks impregnated with whole blood. J ClinMicrobiol. 1991, 29: 1171-1174.
13.
go back to reference Lal AA, Changkasiri S, Hollingdale MR, McCutchan TF: Ribosomal RNA-based diagnosis of Plasmodium falciparum malaria. Mol Biochem Parasitol. 1989, 36: 67-72. 10.1016/0166-6851(89)90201-6.CrossRefPubMed Lal AA, Changkasiri S, Hollingdale MR, McCutchan TF: Ribosomal RNA-based diagnosis of Plasmodium falciparum malaria. Mol Biochem Parasitol. 1989, 36: 67-72. 10.1016/0166-6851(89)90201-6.CrossRefPubMed
14.
go back to reference Singh B, Cox-Singh J, Miller AO, Abdullah MS, Snounou G, Rahman HA: Detection of malaria in Malaysia by nested polymerase chain reaction amplification of dried blood spots on filter papers. Trans R Soc Trop Med Hyg. 1996, 90: 519-521. 10.1016/S0035-9203(96)90302-8.CrossRefPubMed Singh B, Cox-Singh J, Miller AO, Abdullah MS, Snounou G, Rahman HA: Detection of malaria in Malaysia by nested polymerase chain reaction amplification of dried blood spots on filter papers. Trans R Soc Trop Med Hyg. 1996, 90: 519-521. 10.1016/S0035-9203(96)90302-8.CrossRefPubMed
15.
go back to reference Snounou G, Pinheiro L, Gonçalves A, Fonseca L, Dias F, Brown KN, Do Rosario V: The importance of sensitive detection of malaria parasites in the human and insect hosts in epidemiological studies, as shown by the analysis of field samples from Guinea Bissau. Trans R Soc Trop Med Hyg. 1993, 87: 649-653. 10.1016/0035-9203(93)90274-T.CrossRefPubMed Snounou G, Pinheiro L, Gonçalves A, Fonseca L, Dias F, Brown KN, Do Rosario V: The importance of sensitive detection of malaria parasites in the human and insect hosts in epidemiological studies, as shown by the analysis of field samples from Guinea Bissau. Trans R Soc Trop Med Hyg. 1993, 87: 649-653. 10.1016/0035-9203(93)90274-T.CrossRefPubMed
16.
go back to reference Snounou G, Viriyakosol S, Zhu XP, Jarra W, Thaithong S, Brown KN: Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of high prevalence of mixed infections. Mol Biochem Parasitol. 1993, 58: 283-292. 10.1016/0166-6851(93)90050-8.CrossRefPubMed Snounou G, Viriyakosol S, Zhu XP, Jarra W, Thaithong S, Brown KN: Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of high prevalence of mixed infections. Mol Biochem Parasitol. 1993, 58: 283-292. 10.1016/0166-6851(93)90050-8.CrossRefPubMed
17.
go back to reference Lee MA, Tan CH, Aw LT, Tang CS, Singh M, Lee SH, Chia HP, Yap EP: Real time fluorescence-based PCR for detection of malaria parasites. J Clin Microbiol. 2002, 40: 4343-4345. 10.1128/JCM.40.11.4343-4345.2002.PubMedCentralCrossRefPubMed Lee MA, Tan CH, Aw LT, Tang CS, Singh M, Lee SH, Chia HP, Yap EP: Real time fluorescence-based PCR for detection of malaria parasites. J Clin Microbiol. 2002, 40: 4343-4345. 10.1128/JCM.40.11.4343-4345.2002.PubMedCentralCrossRefPubMed
18.
go back to reference Farcas GA, Zhong KJ, Mazzulli T, Kain KC: Evaluation of the RealArt Malaria LC real-time PCR assay for malaria diagnosis. J Clin Microbiol. 2004, 42: 636-638. 10.1128/JCM.42.2.636-638.2004.PubMedCentralCrossRefPubMed Farcas GA, Zhong KJ, Mazzulli T, Kain KC: Evaluation of the RealArt Malaria LC real-time PCR assay for malaria diagnosis. J Clin Microbiol. 2004, 42: 636-638. 10.1128/JCM.42.2.636-638.2004.PubMedCentralCrossRefPubMed
19.
go back to reference Shokoples SE, Ndao M, Kowalewska-Grochowska K, Yanow SK: Multiplex real-time PCR assay for discrimination of Plasmodium species with improved sensitivity for mixed infections. J Clin Microbiol. 2009, 47 (Suppl 4): 975-980.PubMedCentralCrossRefPubMed Shokoples SE, Ndao M, Kowalewska-Grochowska K, Yanow SK: Multiplex real-time PCR assay for discrimination of Plasmodium species with improved sensitivity for mixed infections. J Clin Microbiol. 2009, 47 (Suppl 4): 975-980.PubMedCentralCrossRefPubMed
20.
go back to reference Erdman LK, Kain KC: Molecular diagnostic and surveillance tools for global malaria control. Travel Med Infect Dis. 2008, 6: 82-99. 10.1016/j.tmaid.2007.10.001.CrossRefPubMed Erdman LK, Kain KC: Molecular diagnostic and surveillance tools for global malaria control. Travel Med Infect Dis. 2008, 6: 82-99. 10.1016/j.tmaid.2007.10.001.CrossRefPubMed
21.
go back to reference Luz K, Vasquez D, Zalis MG, Daniel-Ribeiro CT, Alecrim W, Ferreira-da-Cruz MF: Standardization of a very specific and sensitive single PCR for detection of Plasmodium vivax in low parasitized individuals and its usefulness for screening blood donors. Parasitol Res. 2006, 98: 519-524. 10.1007/s00436-005-0085-8.CrossRef Luz K, Vasquez D, Zalis MG, Daniel-Ribeiro CT, Alecrim W, Ferreira-da-Cruz MF: Standardization of a very specific and sensitive single PCR for detection of Plasmodium vivax in low parasitized individuals and its usefulness for screening blood donors. Parasitol Res. 2006, 98: 519-524. 10.1007/s00436-005-0085-8.CrossRef
22.
go back to reference Rubio JM, Benito A, Roche J, Berzosa PJ, Garcia ML, Mico M, Edu M, Alvar J: Semi-nested, multiplex polymerase chain reaction for detection of human malaria parasites and evidence of Plasmodium vivax infection in Equatorial Guinea. Am J Trop Med Hyg. 1999, 60: 183-187.PubMed Rubio JM, Benito A, Roche J, Berzosa PJ, Garcia ML, Mico M, Edu M, Alvar J: Semi-nested, multiplex polymerase chain reaction for detection of human malaria parasites and evidence of Plasmodium vivax infection in Equatorial Guinea. Am J Trop Med Hyg. 1999, 60: 183-187.PubMed
23.
go back to reference Bustin SA: Why the need for qPCR publication guidelines?–The case for MIQE. Methods. 2010, 50: 217-226. 10.1016/j.ymeth.2009.12.006.CrossRefPubMed Bustin SA: Why the need for qPCR publication guidelines?–The case for MIQE. Methods. 2010, 50: 217-226. 10.1016/j.ymeth.2009.12.006.CrossRefPubMed
24.
go back to reference Landis JR, Koch GG: An application of hierarchical kappa-type statistics in the assessment of majority agreement among multiple observers. Biometrics. 1977, 33: 363-374. 10.2307/2529786.CrossRefPubMed Landis JR, Koch GG: An application of hierarchical kappa-type statistics in the assessment of majority agreement among multiple observers. Biometrics. 1977, 33: 363-374. 10.2307/2529786.CrossRefPubMed
25.
go back to reference Graves PM, Richards FO, Ngondi J, Emerson PM, Shargie EB, Endeshaw T, Ceccato P, Ejigsemahu Y, Mosher AW, Hailemariam A, Zerihun M, Teferi T, Ayele B, Mesele A, Yohannes G, Tilahun A, Gebre T: Individual, household, and environmental risk factors for malaria infection in Amhara, Oromia and SNNP regions of Ethiopia. Trans R Soc Trop Med Hyg. 2009, 103: 1211-1220. 10.1016/j.trstmh.2008.11.016.CrossRefPubMed Graves PM, Richards FO, Ngondi J, Emerson PM, Shargie EB, Endeshaw T, Ceccato P, Ejigsemahu Y, Mosher AW, Hailemariam A, Zerihun M, Teferi T, Ayele B, Mesele A, Yohannes G, Tilahun A, Gebre T: Individual, household, and environmental risk factors for malaria infection in Amhara, Oromia and SNNP regions of Ethiopia. Trans R Soc Trop Med Hyg. 2009, 103: 1211-1220. 10.1016/j.trstmh.2008.11.016.CrossRefPubMed
26.
go back to reference Vinetz JM, Gilman RH: Asymptomatic Plasmodium parasitemia and the ecology of malaria transmission. Am J Trop Med Hyg. 2002, 66: 639-640.PubMed Vinetz JM, Gilman RH: Asymptomatic Plasmodium parasitemia and the ecology of malaria transmission. Am J Trop Med Hyg. 2002, 66: 639-640.PubMed
27.
go back to reference Alves FP, Gil LHS, Marrelli MT, Ribolla PEM, Camargo EP, Silva LHP: Asymptomatic carriers of Plasmodium spp. as infection source for malaria vector mosquitoes in the Brazilian amazon. J Med Entomol. 2005, 42: 777-779. 10.1603/0022-2585(2005)042[0777:ACOPSA]2.0.CO;2.CrossRefPubMed Alves FP, Gil LHS, Marrelli MT, Ribolla PEM, Camargo EP, Silva LHP: Asymptomatic carriers of Plasmodium spp. as infection source for malaria vector mosquitoes in the Brazilian amazon. J Med Entomol. 2005, 42: 777-779. 10.1603/0022-2585(2005)042[0777:ACOPSA]2.0.CO;2.CrossRefPubMed
28.
go back to reference Gama BE, Silva-Pires F, Lopes M, Cardoso MA, Britto C, Torres KL, Lima LM, Souza JM, Daniel-Ribeiro CT, Ferreira-da-Cruz MF: Real-time PCR versus conventional PCR for malaria parasite detection in low-grade parasitemia. ExpParasitol. 2007, 116: 427-432. Gama BE, Silva-Pires F, Lopes M, Cardoso MA, Britto C, Torres KL, Lima LM, Souza JM, Daniel-Ribeiro CT, Ferreira-da-Cruz MF: Real-time PCR versus conventional PCR for malaria parasite detection in low-grade parasitemia. ExpParasitol. 2007, 116: 427-432.
29.
go back to reference Payne D: Use and limitations of light microscopy for diagnosing malaria at the primary health care level. Bull World Health Organ. 1988, 66: 621-626.PubMedCentralPubMed Payne D: Use and limitations of light microscopy for diagnosing malaria at the primary health care level. Bull World Health Organ. 1988, 66: 621-626.PubMedCentralPubMed
30.
go back to reference Harris I, Sharrock WW, Bain LM, Gray KA, Bobogare A, Boaz L, Lilley K, Krause D, Vallely A, Johnson ML, Gatton ML, Shanks GD, Cheng Q: A large proportion of asymptomatic Plasmodium infections with low and sub-microscopic parasite densities in the low transmission setting of Temotu Province, Solomon Islands: challenges for malaria diagnostics in an elimination setting. Malar J. 2010, 9: 254-10.1186/1475-2875-9-254.PubMedCentralCrossRefPubMed Harris I, Sharrock WW, Bain LM, Gray KA, Bobogare A, Boaz L, Lilley K, Krause D, Vallely A, Johnson ML, Gatton ML, Shanks GD, Cheng Q: A large proportion of asymptomatic Plasmodium infections with low and sub-microscopic parasite densities in the low transmission setting of Temotu Province, Solomon Islands: challenges for malaria diagnostics in an elimination setting. Malar J. 2010, 9: 254-10.1186/1475-2875-9-254.PubMedCentralCrossRefPubMed
31.
go back to reference Okell LC, Ghani AC, Lyons E, Drakeley CJ: Submicroscopic infection in Plasmodium falciparum-endemic populations: A systematic review and meta-analysis. J Infect Dis. 2009, 200: 1509-1517. 10.1086/644781.CrossRefPubMed Okell LC, Ghani AC, Lyons E, Drakeley CJ: Submicroscopic infection in Plasmodium falciparum-endemic populations: A systematic review and meta-analysis. J Infect Dis. 2009, 200: 1509-1517. 10.1086/644781.CrossRefPubMed
32.
go back to reference Emerson PM, Ngondi J, Biru E, Graves PM, Ejigsemahu Y, Gebre T, Endeshaw T, Genet A, Mosher AW, Zerihun M, Messele A, Richards FO: Integrating an NTD with one of “the big three”: combined malaria and trachoma survey in Amhara Region of Ethiopia. PLoSNegl Trop Dis. 2008, 2: 197-10.1371/journal.pntd.0000197.CrossRef Emerson PM, Ngondi J, Biru E, Graves PM, Ejigsemahu Y, Gebre T, Endeshaw T, Genet A, Mosher AW, Zerihun M, Messele A, Richards FO: Integrating an NTD with one of “the big three”: combined malaria and trachoma survey in Amhara Region of Ethiopia. PLoSNegl Trop Dis. 2008, 2: 197-10.1371/journal.pntd.0000197.CrossRef
33.
go back to reference Alemu A, Tsegaye W, Golassa L, Abebel G: Urban malaria and associated risk factors in Jimma town, south-west Ethiopia. Malar J. 2011, 10: 173-10.1186/1475-2875-10-173.PubMedCentralCrossRefPubMed Alemu A, Tsegaye W, Golassa L, Abebel G: Urban malaria and associated risk factors in Jimma town, south-west Ethiopia. Malar J. 2011, 10: 173-10.1186/1475-2875-10-173.PubMedCentralCrossRefPubMed
34.
go back to reference Shargie EB, Gebre T, Ngondi J, Graves PM, Mosher AW, Emerson PM, Ejigsemahu Y, Endeshaw T, Olana D, WeldeMeskel A, Teferra A, Tadesse Z, Tilahun , Yohannes G, Richards FOJ: Malaria prevalence and mosquito net coverage in Oromia and SNNPR regions of Ethiopia. BMC Publ Health. 2008, 8: 321-10.1186/1471-2458-8-321.CrossRef Shargie EB, Gebre T, Ngondi J, Graves PM, Mosher AW, Emerson PM, Ejigsemahu Y, Endeshaw T, Olana D, WeldeMeskel A, Teferra A, Tadesse Z, Tilahun , Yohannes G, Richards FOJ: Malaria prevalence and mosquito net coverage in Oromia and SNNPR regions of Ethiopia. BMC Publ Health. 2008, 8: 321-10.1186/1471-2458-8-321.CrossRef
35.
go back to reference Mula P, Fernández-Martínez A, de Lucio A, Ramos JM, Reyes F, González V, Benito A, Berzosa P: Detection of high levels of mutations involved in anti-malarial drug resistance in Plasmodium falciparum and Plasmodium vivax at a rural hospital in southern Ethiopia. Malar J. 2011, 10: 214-10.1186/1475-2875-10-214.PubMedCentralCrossRefPubMed Mula P, Fernández-Martínez A, de Lucio A, Ramos JM, Reyes F, González V, Benito A, Berzosa P: Detection of high levels of mutations involved in anti-malarial drug resistance in Plasmodium falciparum and Plasmodium vivax at a rural hospital in southern Ethiopia. Malar J. 2011, 10: 214-10.1186/1475-2875-10-214.PubMedCentralCrossRefPubMed
36.
go back to reference Ashton RA, Kefyalew T, Tesfeye G, Pullan RL, Yadete D, Reithinger R, Kolaczinski JH, Brooker S: School-based surveys of malaria in Oromia Regional State, Ethiopia: a rapid survey method for malaria in low transmission settings. Malar J. 2011, 10: 25-10.1186/1475-2875-10-25.PubMedCentralCrossRefPubMed Ashton RA, Kefyalew T, Tesfeye G, Pullan RL, Yadete D, Reithinger R, Kolaczinski JH, Brooker S: School-based surveys of malaria in Oromia Regional State, Ethiopia: a rapid survey method for malaria in low transmission settings. Malar J. 2011, 10: 25-10.1186/1475-2875-10-25.PubMedCentralCrossRefPubMed
37.
go back to reference Abeku TA, van Oortmarssen GJ, Borsboom G, De Vlas SJ, Habbema JD: Spatial and temporal variations of malaria epidemic risk in Ethiopia: factors involved and implications. Acta Trop. 2003, 87: 331-340. 10.1016/S0001-706X(03)00123-2.CrossRefPubMed Abeku TA, van Oortmarssen GJ, Borsboom G, De Vlas SJ, Habbema JD: Spatial and temporal variations of malaria epidemic risk in Ethiopia: factors involved and implications. Acta Trop. 2003, 87: 331-340. 10.1016/S0001-706X(03)00123-2.CrossRefPubMed
38.
go back to reference Yeshiwondim A, Gopal S, Hailemariam AT, Dengela DO, Patel HP: Spatial analysis of malaria incidence at the village level in areas with unstable transmission in Ethiopia. Int J Health Geogr. 2009, 8: 5-10.1186/1476-072X-8-5.PubMedCentralCrossRefPubMed Yeshiwondim A, Gopal S, Hailemariam AT, Dengela DO, Patel HP: Spatial analysis of malaria incidence at the village level in areas with unstable transmission in Ethiopia. Int J Health Geogr. 2009, 8: 5-10.1186/1476-072X-8-5.PubMedCentralCrossRefPubMed
39.
go back to reference Kiszewski AE, Tekelehaimanot A: A review of the clinical and epidemiologic burdens of epidemic malaria. Am J Trop Med Hyg. 2004, 71 (Suppl 2): 128-135.PubMed Kiszewski AE, Tekelehaimanot A: A review of the clinical and epidemiologic burdens of epidemic malaria. Am J Trop Med Hyg. 2004, 71 (Suppl 2): 128-135.PubMed
40.
go back to reference Smith DL, Guerra CA, Snow RW, Hay SI: Standardizing estimates of the Plasmodium falciparum parasite rate. Malar J. 2007, 6: 131-10.1186/1475-2875-6-131.PubMedCentralCrossRefPubMed Smith DL, Guerra CA, Snow RW, Hay SI: Standardizing estimates of the Plasmodium falciparum parasite rate. Malar J. 2007, 6: 131-10.1186/1475-2875-6-131.PubMedCentralCrossRefPubMed
41.
go back to reference Giha HA, Rosthoj S, Dodoo D, Hviid L, Satti GM, Scheike T, Arnot DE: The epidemiology of febrile malaria episodes in an area of unstable and seasonal transmission. Trans R Soc Trop Med Hyg. 2003, 94: 645-651.CrossRef Giha HA, Rosthoj S, Dodoo D, Hviid L, Satti GM, Scheike T, Arnot DE: The epidemiology of febrile malaria episodes in an area of unstable and seasonal transmission. Trans R Soc Trop Med Hyg. 2003, 94: 645-651.CrossRef
42.
go back to reference Oesterholt MJ, Bousema JT, Mwerinde OK, Harris C, Lushino P, Masokoto A, Mwerinde H, Mosha FW, Drakeley CJ: Spatial and temporal variation in malaria transmission in a low endemicity area in northern Tanzania. Malar J. 2006, 5: 98-10.1186/1475-2875-5-98.PubMedCentralCrossRefPubMed Oesterholt MJ, Bousema JT, Mwerinde OK, Harris C, Lushino P, Masokoto A, Mwerinde H, Mosha FW, Drakeley CJ: Spatial and temporal variation in malaria transmission in a low endemicity area in northern Tanzania. Malar J. 2006, 5: 98-10.1186/1475-2875-5-98.PubMedCentralCrossRefPubMed
43.
go back to reference WHO: Report on the malaria conference in equatorial Africa. Held under the joint auspices of the World Health Organization and of the commission for technical co-operation in Africa south of the Sahara.27 November–9 December, 1950. 1951, Kampala, Uganda. Geneva: World Health Organ Tech Rep Ser, 38:72 WHO: Report on the malaria conference in equatorial Africa. Held under the joint auspices of the World Health Organization and of the commission for technical co-operation in Africa south of the Sahara.27 November–9 December, 1950. 1951, Kampala, Uganda. Geneva: World Health Organ Tech Rep Ser, 38:72
Metadata
Title
Microscopy and molecular biology for the diagnosis and evaluation of malaria in a hospital in a rural area of Ethiopia
Authors
Maria A Santana-Morales
Raquel N Afonso-Lehmann
Maria A Quispe
Francisco Reyes
Pedro Berzosa
Agustin Benito
Basilio Valladares
Enrique Martinez-Carretero
Publication date
01-12-2012
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2012
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/1475-2875-11-199

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