A direct comparison of real time PCR on plasma and blood to detect Plasmodium falciparum infection in children

Abigail A. Lamikanra; Carlota Dobão; Alfons Jiménez; Augusto Nhabomba; Hoi P. Tsang; Caterina Guinovart; Maria N. Manaca; Llorenç Quinto; Ruth Aguilar; Pau Cisterá; Pedro L. Alonso; David J. Roberts; Alfredo Mayor

doi:10.1186/1475-2875-11-201

A direct comparison of real time PCR on plasma and blood to detect Plasmodium falciparum infection in children

Abigail A. Lamikanra^*, Carlota Dobão, Alfons Jiménez, Augusto Nhabomba, Hoi P. Tsang, Caterina Guinovart, Maria N. Manaca, Llorenç Quinto, Ruth Aguilar, Pau Cisterá, Pedro L. Alonso, David J. Roberts, Alfredo Mayor

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

Abstract

Background: Estimation of Plasmodium falciparum parasitaemia can vary with the method used and time of sampling. Quantitative real time PCR (qPCR) on whole blood or plasma samples has previously been shown to be more sensitive than thick film microscopy. However the efficiencies of each method have not been compared using samples obtained from infants less than one year old. Methods: A multiple of statistical approaches were used to compare the performance of qPCR on whole blood or plasma to detect the 18 S ribosomal gene of P. falciparum in 548 samples from children aged 2.5 or 24 months. Parasite prevalence in matched samples was compared using Mcnemars test and agreement of positive results quantified as Kappa scores. Parasite prevalences between different age groups were compared by Fishers test. Results from analyses by thick film microscopy were also available from children at 24 months and their correlation to each qPCR method examined by the Spearmans test. Finally the association of P. falciparum infection with the incidence of multiple malaria episodes from contact to 24 months of age was evaluated using negative binomial regression. Results: These analyses showed that qPCR from whole blood detected approximately 3-fold more cases of infection than plasma qPCR. Both qPCR methods agreed well with each other although qPCR from plasma had a greater agreement with microscopy (96.85%) than did qPCR from blood (69.7%). At 24 months the prevalence of infection detected by all methods was associated with anaemia (p≥0.05). Conclusions: The data presented here demonstrates that low levels of parasitaemia are better detected by qPCR using parasite DNA from whole blood than from plasma. However plasma samples provide a viable substitute when parasite smears are unavailable.

Original language	English
Article number	201
Journal	Malaria Journal
Volume	11
DOIs	https://doi.org/10.1186/1475-2875-11-201
Publication status	Published - 2012
Externally published	Yes

Bibliographical note

Funding Information:
We thank all children and their families for their participation in the study; the field workers, field supervisors, laboratory staff, data managers and other staff at CISM for their work during the study. The study was funded by a EU Framework Program 6 STREP project (Malaria age exposure, Project reference: 18902), the Instituto de Salud Carlos III (Ayuda de incentivación a la participación en proyectos del Espació Europeo de Investigación) and the Spanish Ministerio de Educación y Ciencia (Project reference: A107190024). C.G. was supported by a grant from the Spanish Ministry of Health (CM04/00028), C.D. from the Spanish Ministerio de Ciencia e Innovación (RYC-2008-02631) and A.M. from the Instituto de Salud carlos III (CP-04/00220). A.L. H.P.T and D.J.R. are supported by the National Health Service Blood and Transplant (NHSBT), National Institutes of Health Research (NIHR) Biomedical Research Centres funding scheme and NIHR Programme Grant NIHR-RP-PG-0310-1004-AN. The Manhiça Health Research Centre receives core funding from the Spanish Agency for International Cooperation and Development (AECID).

Keywords

Anaemia
Malaria
Plasma
qPCR

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1186/1475-2875-11-201

Cite this

Lamikanra, A. A., Dobão, C., Jiménez, A., Nhabomba, A., Tsang, H. P., Guinovart, C., Manaca, M. N., Quinto, L., Aguilar, R., Cisterá, P., Alonso, P. L., Roberts, D. J., & Mayor, A. (2012). A direct comparison of real time PCR on plasma and blood to detect Plasmodium falciparum infection in children. Malaria Journal, 11, Article 201. https://doi.org/10.1186/1475-2875-11-201

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title = "A direct comparison of real time PCR on plasma and blood to detect Plasmodium falciparum infection in children",

abstract = "Background: Estimation of Plasmodium falciparum parasitaemia can vary with the method used and time of sampling. Quantitative real time PCR (qPCR) on whole blood or plasma samples has previously been shown to be more sensitive than thick film microscopy. However the efficiencies of each method have not been compared using samples obtained from infants less than one year old. Methods: A multiple of statistical approaches were used to compare the performance of qPCR on whole blood or plasma to detect the 18 S ribosomal gene of P. falciparum in 548 samples from children aged 2.5 or 24 months. Parasite prevalence in matched samples was compared using Mcnemars test and agreement of positive results quantified as Kappa scores. Parasite prevalences between different age groups were compared by Fishers test. Results from analyses by thick film microscopy were also available from children at 24 months and their correlation to each qPCR method examined by the Spearmans test. Finally the association of P. falciparum infection with the incidence of multiple malaria episodes from contact to 24 months of age was evaluated using negative binomial regression. Results: These analyses showed that qPCR from whole blood detected approximately 3-fold more cases of infection than plasma qPCR. Both qPCR methods agreed well with each other although qPCR from plasma had a greater agreement with microscopy (96.85%) than did qPCR from blood (69.7%). At 24 months the prevalence of infection detected by all methods was associated with anaemia (p≥0.05). Conclusions: The data presented here demonstrates that low levels of parasitaemia are better detected by qPCR using parasite DNA from whole blood than from plasma. However plasma samples provide a viable substitute when parasite smears are unavailable.",

keywords = "Anaemia, Malaria, Plasma, qPCR",

author = "Lamikanra, {Abigail A.} and Carlota Dob{\~a}o and Alfons Jim{\'e}nez and Augusto Nhabomba and Tsang, {Hoi P.} and Caterina Guinovart and Manaca, {Maria N.} and Lloren{\c c} Quinto and Ruth Aguilar and Pau Cister{\'a} and Alonso, {Pedro L.} and Roberts, {David J.} and Alfredo Mayor",

note = "Funding Information: We thank all children and their families for their participation in the study; the field workers, field supervisors, laboratory staff, data managers and other staff at CISM for their work during the study. The study was funded by a EU Framework Program 6 STREP project (Malaria age exposure, Project reference: 18902), the Instituto de Salud Carlos III (Ayuda de incentivaci{\'o}n a la participaci{\'o}n en proyectos del Espaci{\'o} Europeo de Investigaci{\'o}n) and the Spanish Ministerio de Educaci{\'o}n y Ciencia (Project reference: A107190024). C.G. was supported by a grant from the Spanish Ministry of Health (CM04/00028), C.D. from the Spanish Ministerio de Ciencia e Innovaci{\'o}n (RYC-2008-02631) and A.M. from the Instituto de Salud carlos III (CP-04/00220). A.L. H.P.T and D.J.R. are supported by the National Health Service Blood and Transplant (NHSBT), National Institutes of Health Research (NIHR) Biomedical Research Centres funding scheme and NIHR Programme Grant NIHR-RP-PG-0310-1004-AN. The Manhi{\c c}a Health Research Centre receives core funding from the Spanish Agency for International Cooperation and Development (AECID).",

year = "2012",

doi = "10.1186/1475-2875-11-201",

language = "English",

volume = "11",

journal = "Malaria Journal",

issn = "1475-2875",

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TY - JOUR

T1 - A direct comparison of real time PCR on plasma and blood to detect Plasmodium falciparum infection in children

AU - Lamikanra, Abigail A.

AU - Dobão, Carlota

AU - Jiménez, Alfons

AU - Nhabomba, Augusto

AU - Tsang, Hoi P.

AU - Guinovart, Caterina

AU - Manaca, Maria N.

AU - Quinto, Llorenç

AU - Aguilar, Ruth

AU - Cisterá, Pau

AU - Alonso, Pedro L.

AU - Roberts, David J.

AU - Mayor, Alfredo

N1 - Funding Information: We thank all children and their families for their participation in the study; the field workers, field supervisors, laboratory staff, data managers and other staff at CISM for their work during the study. The study was funded by a EU Framework Program 6 STREP project (Malaria age exposure, Project reference: 18902), the Instituto de Salud Carlos III (Ayuda de incentivación a la participación en proyectos del Espació Europeo de Investigación) and the Spanish Ministerio de Educación y Ciencia (Project reference: A107190024). C.G. was supported by a grant from the Spanish Ministry of Health (CM04/00028), C.D. from the Spanish Ministerio de Ciencia e Innovación (RYC-2008-02631) and A.M. from the Instituto de Salud carlos III (CP-04/00220). A.L. H.P.T and D.J.R. are supported by the National Health Service Blood and Transplant (NHSBT), National Institutes of Health Research (NIHR) Biomedical Research Centres funding scheme and NIHR Programme Grant NIHR-RP-PG-0310-1004-AN. The Manhiça Health Research Centre receives core funding from the Spanish Agency for International Cooperation and Development (AECID).

PY - 2012

Y1 - 2012

N2 - Background: Estimation of Plasmodium falciparum parasitaemia can vary with the method used and time of sampling. Quantitative real time PCR (qPCR) on whole blood or plasma samples has previously been shown to be more sensitive than thick film microscopy. However the efficiencies of each method have not been compared using samples obtained from infants less than one year old. Methods: A multiple of statistical approaches were used to compare the performance of qPCR on whole blood or plasma to detect the 18 S ribosomal gene of P. falciparum in 548 samples from children aged 2.5 or 24 months. Parasite prevalence in matched samples was compared using Mcnemars test and agreement of positive results quantified as Kappa scores. Parasite prevalences between different age groups were compared by Fishers test. Results from analyses by thick film microscopy were also available from children at 24 months and their correlation to each qPCR method examined by the Spearmans test. Finally the association of P. falciparum infection with the incidence of multiple malaria episodes from contact to 24 months of age was evaluated using negative binomial regression. Results: These analyses showed that qPCR from whole blood detected approximately 3-fold more cases of infection than plasma qPCR. Both qPCR methods agreed well with each other although qPCR from plasma had a greater agreement with microscopy (96.85%) than did qPCR from blood (69.7%). At 24 months the prevalence of infection detected by all methods was associated with anaemia (p≥0.05). Conclusions: The data presented here demonstrates that low levels of parasitaemia are better detected by qPCR using parasite DNA from whole blood than from plasma. However plasma samples provide a viable substitute when parasite smears are unavailable.

AB - Background: Estimation of Plasmodium falciparum parasitaemia can vary with the method used and time of sampling. Quantitative real time PCR (qPCR) on whole blood or plasma samples has previously been shown to be more sensitive than thick film microscopy. However the efficiencies of each method have not been compared using samples obtained from infants less than one year old. Methods: A multiple of statistical approaches were used to compare the performance of qPCR on whole blood or plasma to detect the 18 S ribosomal gene of P. falciparum in 548 samples from children aged 2.5 or 24 months. Parasite prevalence in matched samples was compared using Mcnemars test and agreement of positive results quantified as Kappa scores. Parasite prevalences between different age groups were compared by Fishers test. Results from analyses by thick film microscopy were also available from children at 24 months and their correlation to each qPCR method examined by the Spearmans test. Finally the association of P. falciparum infection with the incidence of multiple malaria episodes from contact to 24 months of age was evaluated using negative binomial regression. Results: These analyses showed that qPCR from whole blood detected approximately 3-fold more cases of infection than plasma qPCR. Both qPCR methods agreed well with each other although qPCR from plasma had a greater agreement with microscopy (96.85%) than did qPCR from blood (69.7%). At 24 months the prevalence of infection detected by all methods was associated with anaemia (p≥0.05). Conclusions: The data presented here demonstrates that low levels of parasitaemia are better detected by qPCR using parasite DNA from whole blood than from plasma. However plasma samples provide a viable substitute when parasite smears are unavailable.

KW - Anaemia

KW - Malaria

KW - Plasma

KW - qPCR

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DO - 10.1186/1475-2875-11-201

M3 - Article

C2 - 22704637

AN - SCOPUS:84862216520

SN - 1475-2875

VL - 11

JO - Malaria Journal

JF - Malaria Journal

M1 - 201

ER -

A direct comparison of real time PCR on plasma and blood to detect Plasmodium falciparum infection in children

Abstract

Bibliographical note

Keywords

UN SDGs

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