Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest Ethiopia

Abstract Background Low malaria parasitaemia is a diagnostic challenge in pregnancy, leading to false negative microscopy and rapid diagnostic test (RDT) results. However, these submicroscopic or subpatent infections could cause adverse pregnancy outcomes. Thus, evaluating the diagnostic performance...

Full description

Saved in:
Bibliographic Details
Main Authors: Zemenu Tamir, Abebe Animut, Sisay Dugassa, Araya Gebresilassie, Mahlet Belachew, Adugna Abera, Berhanu Erko
Format: Article
Language:English
Published: BMC 2025-01-01
Series:Malaria Journal
Subjects:
Online Access:https://doi.org/10.1186/s12936-025-05256-2
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1832586041289080832
author Zemenu Tamir
Abebe Animut
Sisay Dugassa
Araya Gebresilassie
Mahlet Belachew
Adugna Abera
Berhanu Erko
author_facet Zemenu Tamir
Abebe Animut
Sisay Dugassa
Araya Gebresilassie
Mahlet Belachew
Adugna Abera
Berhanu Erko
author_sort Zemenu Tamir
collection DOAJ
description Abstract Background Low malaria parasitaemia is a diagnostic challenge in pregnancy, leading to false negative microscopy and rapid diagnostic test (RDT) results. However, these submicroscopic or subpatent infections could cause adverse pregnancy outcomes. Thus, evaluating the diagnostic performance of microscopy, RDT, and multiplex qPCR in pregnancy is vital for informed decisions. Methods A total of 835 peripheral blood and 372 placental blood samples were collected from 835 pregnant women attending first antenatal care or admitted for delivery at selected health facilities in northwest Ethiopia between November 2021 and July 2022. In multiplex qPCR, all microscopy and/or RDT positive samples were extracted and amplified individually, whereas all samples negative by both RDT and microscopy were extracted after pooling ten samples together and tested for Plasmodium genus. The diagnostic performance of microscopy, RDT, and multiplex qPCR in pregnancy was compared and evaluated against each other. Results Using multiplex qPCR as a reference test, microscopy had a sensitivity of 73.8% (95% confidence interval (CI): 65.9–80.7) and 62.2% (95% CI: 46.5–76.2) to detect Plasmodium parasites in peripheral and placental blood samples, respectively, with a 100% (95% CI: 98.9–100) specificity in both samples. Similarly, the RDT had a sensitivity of 67.6% (95% CI: 59.3–75.1) and a specificity of 96.5% (95% CI: 94.9–97.8) for Plasmodium infection diagnosis in peripheral blood and a sensitivity of 62.2% (95% CI: 46.5–76.2) and a specificity of 98.8% (95% CI: 96.9–99.7) in placental blood samples. Considering microscopy as a reference test, multiplex qPCR showed a sensitivity of 100% (95% CI: 96.6–100) and a specificity of 94.8% (95% CI: 93.0–96.3) to diagnose Plasmodium infections in both peripheral and placental blood samples. Pooled multiplex qPCR detected 34 peripheral and 12 placental blood Plasmodium infections from microscopy and RDT negative samples. The pooled assay obviated about half of the reactions and its testing costs. Microscopy showed almost perfect agreement (κ = 0.823) with multiplex qPCR for detecting malaria parasites in pregnancy, whereas the RDT showed a substantial agreement (κ = 0.684). Conclusion Multiplex qPCR had a better performance for Plasmodium infection diagnosis in pregnancy compared to microscopy and RDT. Pooled multiplex qPCR could be a sensitive and resource-efficient strategy for epidemiological surveillance of Plasmodium infections in pregnancy.
format Article
id doaj-art-281ad03cc42a4957ada691df43348c43
institution Kabale University
issn 1475-2875
language English
publishDate 2025-01-01
publisher BMC
record_format Article
series Malaria Journal
spelling doaj-art-281ad03cc42a4957ada691df43348c432025-01-26T12:15:59ZengBMCMalaria Journal1475-28752025-01-0124111110.1186/s12936-025-05256-2Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest EthiopiaZemenu Tamir0Abebe Animut1Sisay Dugassa2Araya Gebresilassie3Mahlet Belachew4Adugna Abera5Berhanu Erko6Department of Medical Laboratory Sciences, College of Health Sciences, Addis Ababa UniversityAklilu Lemma Institute of Pathobiology, Addis Ababa UniversityAklilu Lemma Institute of Pathobiology, Addis Ababa UniversityDepartment of Zoological Sciences, College of Natural and Computational Sciences, Addis Ababa UniversityMalaria and Neglected Tropical Diseases Research Team, Ethiopian Public Health InstituteMalaria and Neglected Tropical Diseases Research Team, Ethiopian Public Health InstituteAklilu Lemma Institute of Pathobiology, Addis Ababa UniversityAbstract Background Low malaria parasitaemia is a diagnostic challenge in pregnancy, leading to false negative microscopy and rapid diagnostic test (RDT) results. However, these submicroscopic or subpatent infections could cause adverse pregnancy outcomes. Thus, evaluating the diagnostic performance of microscopy, RDT, and multiplex qPCR in pregnancy is vital for informed decisions. Methods A total of 835 peripheral blood and 372 placental blood samples were collected from 835 pregnant women attending first antenatal care or admitted for delivery at selected health facilities in northwest Ethiopia between November 2021 and July 2022. In multiplex qPCR, all microscopy and/or RDT positive samples were extracted and amplified individually, whereas all samples negative by both RDT and microscopy were extracted after pooling ten samples together and tested for Plasmodium genus. The diagnostic performance of microscopy, RDT, and multiplex qPCR in pregnancy was compared and evaluated against each other. Results Using multiplex qPCR as a reference test, microscopy had a sensitivity of 73.8% (95% confidence interval (CI): 65.9–80.7) and 62.2% (95% CI: 46.5–76.2) to detect Plasmodium parasites in peripheral and placental blood samples, respectively, with a 100% (95% CI: 98.9–100) specificity in both samples. Similarly, the RDT had a sensitivity of 67.6% (95% CI: 59.3–75.1) and a specificity of 96.5% (95% CI: 94.9–97.8) for Plasmodium infection diagnosis in peripheral blood and a sensitivity of 62.2% (95% CI: 46.5–76.2) and a specificity of 98.8% (95% CI: 96.9–99.7) in placental blood samples. Considering microscopy as a reference test, multiplex qPCR showed a sensitivity of 100% (95% CI: 96.6–100) and a specificity of 94.8% (95% CI: 93.0–96.3) to diagnose Plasmodium infections in both peripheral and placental blood samples. Pooled multiplex qPCR detected 34 peripheral and 12 placental blood Plasmodium infections from microscopy and RDT negative samples. The pooled assay obviated about half of the reactions and its testing costs. Microscopy showed almost perfect agreement (κ = 0.823) with multiplex qPCR for detecting malaria parasites in pregnancy, whereas the RDT showed a substantial agreement (κ = 0.684). Conclusion Multiplex qPCR had a better performance for Plasmodium infection diagnosis in pregnancy compared to microscopy and RDT. Pooled multiplex qPCR could be a sensitive and resource-efficient strategy for epidemiological surveillance of Plasmodium infections in pregnancy.https://doi.org/10.1186/s12936-025-05256-2Plasmodium infectionMicroscopyMultiplex real-time PCRRapid diagnostic testsDiagnostic performance
spellingShingle Zemenu Tamir
Abebe Animut
Sisay Dugassa
Araya Gebresilassie
Mahlet Belachew
Adugna Abera
Berhanu Erko
Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest Ethiopia
Malaria Journal
Plasmodium infection
Microscopy
Multiplex real-time PCR
Rapid diagnostic tests
Diagnostic performance
title Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest Ethiopia
title_full Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest Ethiopia
title_fullStr Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest Ethiopia
title_full_unstemmed Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest Ethiopia
title_short Comparative performance of microscopy, rapid diagnostic tests, and multiplex real-time PCR for detection of malaria parasites among pregnant women in northwest Ethiopia
title_sort comparative performance of microscopy rapid diagnostic tests and multiplex real time pcr for detection of malaria parasites among pregnant women in northwest ethiopia
topic Plasmodium infection
Microscopy
Multiplex real-time PCR
Rapid diagnostic tests
Diagnostic performance
url https://doi.org/10.1186/s12936-025-05256-2
work_keys_str_mv AT zemenutamir comparativeperformanceofmicroscopyrapiddiagnostictestsandmultiplexrealtimepcrfordetectionofmalariaparasitesamongpregnantwomeninnorthwestethiopia
AT abebeanimut comparativeperformanceofmicroscopyrapiddiagnostictestsandmultiplexrealtimepcrfordetectionofmalariaparasitesamongpregnantwomeninnorthwestethiopia
AT sisaydugassa comparativeperformanceofmicroscopyrapiddiagnostictestsandmultiplexrealtimepcrfordetectionofmalariaparasitesamongpregnantwomeninnorthwestethiopia
AT arayagebresilassie comparativeperformanceofmicroscopyrapiddiagnostictestsandmultiplexrealtimepcrfordetectionofmalariaparasitesamongpregnantwomeninnorthwestethiopia
AT mahletbelachew comparativeperformanceofmicroscopyrapiddiagnostictestsandmultiplexrealtimepcrfordetectionofmalariaparasitesamongpregnantwomeninnorthwestethiopia
AT adugnaabera comparativeperformanceofmicroscopyrapiddiagnostictestsandmultiplexrealtimepcrfordetectionofmalariaparasitesamongpregnantwomeninnorthwestethiopia
AT berhanuerko comparativeperformanceofmicroscopyrapiddiagnostictestsandmultiplexrealtimepcrfordetectionofmalariaparasitesamongpregnantwomeninnorthwestethiopia