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Published in: Malaria Journal 1/2014

Open Access 01-12-2014 | Methodology

Limit of blank and limit of detection of Plasmodium falciparum thick blood smear microscopy in a routine setting in Central Africa

Authors: Fanny Joanny, Sascha JZ Löhr, Thomas Engleitner, Bertrand Lell, Benjamin Mordmüller

Published in: Malaria Journal | Issue 1/2014

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Abstract

Background

Proper malaria diagnosis depends on the detection of asexual forms of Plasmodium spp. in the blood. Thick blood smear microscopy is the accepted gold standard of malaria diagnosis and is widely implemented. Surprisingly, diagnostic performance of this method is not well investigated and many clinicians in African routine settings base treatment decisions independent of microscopy results. This leads to overtreatment and poor management of other febrile diseases. Implementation of quality control programmes is recommended, but requires sustained funding, external logistic support and constant training and supervision of the staff. This study describes an easily applicable method to assess the performance of thick blood smear microscopy by determining the limit of blank and limit of detection. These two values are representative of the diagnostic quality and allow the correct discrimination between positive and negative samples.

Methods

Standard-conform methodology was applied and adapted to determine the limit of blank and the limit of detection of two thick blood smear microscopy methods (WHO and Lambaréné method) in a research centre in Lambaréné, Gabon. Duplicates of negative and low parasitaemia thick blood smears were read by several microscopists. The mean and standard deviation of the results were used to calculate the limit of blank and subsequently the limit of detection.

Results

The limit of blank was 0 parasites/μL for both methods. The limit of detection was 62 and 88 parasites/μL for the Lambaréné and WHO method, respectively.

Conclusion

With a simple, back-of-the-envelope calculation, the performance of two malaria microscopy methods can be measured. These results are specific for each diagnostic unit and cannot be generalized but implementation of a system to control microscopy performance can improve confidence in parasitological results and thereby strengthen malaria control.
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Literature
1.
go back to reference WHO: World Malaria Report 2012. r, Geneva: World Health Organization WHO: World Malaria Report 2012. r, Geneva: World Health Organization
2.
go back to reference Murray CJ, Rosenfeld LC, Lim SS, Andrews KG, Foreman KJ, Haring D, Fullman N, Naghavi M, Lozano R, Lopez AD: Global malaria mortality between 1980 and 2010: a systematic analysis. Lancet. 2012, 379: 413-431. 10.1016/S0140-6736(12)60034-8.CrossRefPubMed Murray CJ, Rosenfeld LC, Lim SS, Andrews KG, Foreman KJ, Haring D, Fullman N, Naghavi M, Lozano R, Lopez AD: Global malaria mortality between 1980 and 2010: a systematic analysis. Lancet. 2012, 379: 413-431. 10.1016/S0140-6736(12)60034-8.CrossRefPubMed
3.
go back to reference Long EG: Requirements for diagnosis of malaria at different levels of the laboratory network in Africa. Am J Clin Pathol. 2009, 131: 858-860. 10.1309/AJCPVX71BXWOVWBY.CrossRefPubMed Long EG: Requirements for diagnosis of malaria at different levels of the laboratory network in Africa. Am J Clin Pathol. 2009, 131: 858-860. 10.1309/AJCPVX71BXWOVWBY.CrossRefPubMed
4.
go back to reference Mordmüller B, Kremsner PG: Malarial parasites vs. antimalarials: never-ending rumble in the jungle. Curr Mol Med. 2006, 6: 247-251. 10.2174/156652406776055122.CrossRefPubMed Mordmüller B, Kremsner PG: Malarial parasites vs. antimalarials: never-ending rumble in the jungle. Curr Mol Med. 2006, 6: 247-251. 10.2174/156652406776055122.CrossRefPubMed
5.
go back to reference Giemsa G: Eine Vereinfachung und Vervollkommnung meiner Methylenblau-Eosin-Färbemethode zur Erzielung der Romanowsky-Nocht’schen Chromatinfärbung. Zentralbl Bakteriol A. 1904, 32: 307-313. Giemsa G: Eine Vereinfachung und Vervollkommnung meiner Methylenblau-Eosin-Färbemethode zur Erzielung der Romanowsky-Nocht’schen Chromatinfärbung. Zentralbl Bakteriol A. 1904, 32: 307-313.
6.
go back to reference Aregawi M, Cibulskis RE, Lynch M, Williams R, World Health Organization, Global Malaria Programme: World Malaria Report 2011. 2011, Geneva: World Health Organization Aregawi M, Cibulskis RE, Lynch M, Williams R, World Health Organization, Global Malaria Programme: World Malaria Report 2011. 2011, Geneva: World Health Organization
7.
go back to reference Metzger WG, Mordmüller BG, Kremsner PG: Malaria pigment in leucocytes. Trans R Soc Trop Med Hyg. 1995, 89: 637-638. 10.1016/0035-9203(95)90423-9.CrossRefPubMed Metzger WG, Mordmüller BG, Kremsner PG: Malaria pigment in leucocytes. Trans R Soc Trop Med Hyg. 1995, 89: 637-638. 10.1016/0035-9203(95)90423-9.CrossRefPubMed
8.
go back to reference Planche T, Krishna S, Kombila M, Engel K, Faucher JF, Ngou-Milama E, Kremsner PG: Comparison of methods for the rapid laboratory assessment of children with malaria. Am J Trop Med Hyg. 2001, 65: 599-602.PubMed Planche T, Krishna S, Kombila M, Engel K, Faucher JF, Ngou-Milama E, Kremsner PG: Comparison of methods for the rapid laboratory assessment of children with malaria. Am J Trop Med Hyg. 2001, 65: 599-602.PubMed
9.
go back to reference Lell B, Missinou MA, Issifou S, Matsiegui P-B, Olola CHO, Taylor TE, Kremsner PG: Assessment of a simplified method for counting leukocytic malaria pigment. Am J Trop Med Hyg. 2005, 73: 588-592.PubMed Lell B, Missinou MA, Issifou S, Matsiegui P-B, Olola CHO, Taylor TE, Kremsner PG: Assessment of a simplified method for counting leukocytic malaria pigment. Am J Trop Med Hyg. 2005, 73: 588-592.PubMed
10.
go back to reference Barat L, Chipipa J, Kolczak M, Sukwa T: Does the availability of blood slide microscopy for malaria at health centers improve the management of persons with fever in Zambia?. Am J Trop Med Hyg. 1999, 60: 1024-1030.PubMed Barat L, Chipipa J, Kolczak M, Sukwa T: Does the availability of blood slide microscopy for malaria at health centers improve the management of persons with fever in Zambia?. Am J Trop Med Hyg. 1999, 60: 1024-1030.PubMed
11.
go back to reference Zurovac D, Midia B, Ochola SA, English M, Snow RW: Microscopy and outpatient malaria case management among older children and adults in Kenya. Trop Med Int Health. 2006, 11: 432-440. 10.1111/j.1365-3156.2006.01587.x.CrossRefPubMed Zurovac D, Midia B, Ochola SA, English M, Snow RW: Microscopy and outpatient malaria case management among older children and adults in Kenya. Trop Med Int Health. 2006, 11: 432-440. 10.1111/j.1365-3156.2006.01587.x.CrossRefPubMed
12.
go back to reference Ngasala B, Mubi M, Warsame M, Petzold MG, Massele AY, Gustafsson LL, Tomson G, Premji Z, Bjorkman A: Impact of training in clinical and microscopy diagnosis of childhood malaria on antimalarial drug prescription and health outcome at primary health care level in Tanzania: a randomized controlled trial. Malar J. 2008, 7: 199-10.1186/1475-2875-7-199.PubMedCentralCrossRefPubMed Ngasala B, Mubi M, Warsame M, Petzold MG, Massele AY, Gustafsson LL, Tomson G, Premji Z, Bjorkman A: Impact of training in clinical and microscopy diagnosis of childhood malaria on antimalarial drug prescription and health outcome at primary health care level in Tanzania: a randomized controlled trial. Malar J. 2008, 7: 199-10.1186/1475-2875-7-199.PubMedCentralCrossRefPubMed
13.
go back to reference Mukadi P, Gillet P, Lukuka A, Atua B, Kahodi S, Lokombe J, Muyembe J-J, Jacobs J: External quality assessment of malaria microscopy in the Democratic Republic of the Congo. Malar J. 2011, 10: 308-10.1186/1475-2875-10-308.PubMedCentralCrossRefPubMed Mukadi P, Gillet P, Lukuka A, Atua B, Kahodi S, Lokombe J, Muyembe J-J, Jacobs J: External quality assessment of malaria microscopy in the Democratic Republic of the Congo. Malar J. 2011, 10: 308-10.1186/1475-2875-10-308.PubMedCentralCrossRefPubMed
14.
go back to reference Tholen DW: Protocols for Determination of Limits of Detection and Limits of Quantitation: Approved Guideline EP17-A. 2004, Wayne, Pa: NCCLS Tholen DW: Protocols for Determination of Limits of Detection and Limits of Quantitation: Approved Guideline EP17-A. 2004, Wayne, Pa: NCCLS
15.
go back to reference Linnet K: Nonparametric estimation of reference intervals by simple and bootstrap-based procedures. Clin Chem. 2000, 46 (6 Pt 1): 867-869.PubMed Linnet K: Nonparametric estimation of reference intervals by simple and bootstrap-based procedures. Clin Chem. 2000, 46 (6 Pt 1): 867-869.PubMed
16.
go back to reference Wiehart J: Proofs of the distribution law of the second order moment statistics. Biometrika. 1948, 35 (Pts 1–2): 55-57.CrossRefPubMed Wiehart J: Proofs of the distribution law of the second order moment statistics. Biometrika. 1948, 35 (Pts 1–2): 55-57.CrossRefPubMed
17.
go back to reference Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA: A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg. 1999, 60: 687-692.PubMed Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA: A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg. 1999, 60: 687-692.PubMed
18.
go back to reference Jorgensen E, Pedersen AR: How to obtain those nasty standard errors from transformed data – and why they should not be used. Aarhus Univ Det Jordbrugsvidenskabelige Fak. 1998, 7: 1-20. Jorgensen E, Pedersen AR: How to obtain those nasty standard errors from transformed data – and why they should not be used. Aarhus Univ Det Jordbrugsvidenskabelige Fak. 1998, 7: 1-20.
19.
go back to reference World Health Organization: Basic Malaria Microscopy. 2010, Geneva: WHO World Health Organization: Basic Malaria Microscopy. 2010, Geneva: WHO
20.
go back to reference Ochola LB, Vounatsou P, Smith T, Mabaso MLH, Newton CRJC: 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 MLH, Newton CRJC: 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
21.
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
22.
go back to reference Bruce-Chwatt LJ: From Laveran’s discovery to DNA probes: new trends in diagnosis of malaria. Lancet. 1987, 2: 1509-1511.CrossRefPubMed Bruce-Chwatt LJ: From Laveran’s discovery to DNA probes: new trends in diagnosis of malaria. Lancet. 1987, 2: 1509-1511.CrossRefPubMed
24.
go back to reference Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH: A review of malaria diagnostic tools: microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg. 2007, 77 (6 Suppl): 119-127.PubMed Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH: A review of malaria diagnostic tools: microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg. 2007, 77 (6 Suppl): 119-127.PubMed
25.
go back to reference Zurovac D, Larson BA, Akhwale W, Snow RW: The financial and clinical implications of adult malaria diagnosis using microscopy in Kenya. Trop Med Int Health. 2006, 11: 1185-1194. 10.1111/j.1365-3156.2006.01674.x.CrossRefPubMed Zurovac D, Larson BA, Akhwale W, Snow RW: The financial and clinical implications of adult malaria diagnosis using microscopy in Kenya. Trop Med Int Health. 2006, 11: 1185-1194. 10.1111/j.1365-3156.2006.01674.x.CrossRefPubMed
26.
go back to reference McElroy PD, Beier JC, Oster CN, Beadle C, Sherwood JA, Oloo AJ, Hoffman SL: Predicting outcome in malaria: correlation between rate of exposure to infected mosquitoes and level of Plasmodium falciparum parasitemia. Am J Trop Med Hyg. 1994, 51: 523-532.PubMed McElroy PD, Beier JC, Oster CN, Beadle C, Sherwood JA, Oloo AJ, Hoffman SL: Predicting outcome in malaria: correlation between rate of exposure to infected mosquitoes and level of Plasmodium falciparum parasitemia. Am J Trop Med Hyg. 1994, 51: 523-532.PubMed
27.
go back to reference Moorthy V, Reed Z, Smith PG: Measurement of malaria vaccine efficacy in phase III trials: report of a WHO consultation. Vaccine. 2007, 25: 5115-5123. 10.1016/j.vaccine.2007.01.085.CrossRefPubMed Moorthy V, Reed Z, Smith PG: Measurement of malaria vaccine efficacy in phase III trials: report of a WHO consultation. Vaccine. 2007, 25: 5115-5123. 10.1016/j.vaccine.2007.01.085.CrossRefPubMed
28.
go back to reference Lillie PJ, Duncan CJA, Sheehy SH, Meyer J, O’Hara GA, Gilbert SC, Hill AVS: Distinguishing malaria and influenza: early clinical features in controlled human experimental infection studies. Travel Med Infect Dis. 2012, 10: 192-196. 10.1016/j.tmaid.2012.03.008.PubMedCentralCrossRefPubMed Lillie PJ, Duncan CJA, Sheehy SH, Meyer J, O’Hara GA, Gilbert SC, Hill AVS: Distinguishing malaria and influenza: early clinical features in controlled human experimental infection studies. Travel Med Infect Dis. 2012, 10: 192-196. 10.1016/j.tmaid.2012.03.008.PubMedCentralCrossRefPubMed
Metadata
Title
Limit of blank and limit of detection of Plasmodium falciparum thick blood smear microscopy in a routine setting in Central Africa
Authors
Fanny Joanny
Sascha JZ Löhr
Thomas Engleitner
Bertrand Lell
Benjamin Mordmüller
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2014
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/1475-2875-13-234

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