Malaria prevalence defined by microscopy, antigen detection, DNA amplification and total nucleic acid amplification in a malaria‐endemic region during the peak malaria transmission season

Waitumbi, John N.; Gerlach, Jay; Afonina, Irina; Anyona, Samuel B.; Koros, Joseph N.; Siangla, Joram; Ankoudinova, Irina; Singhal, Mitra C.; Watts, Kate; Polhemus, Mark E.; Vermeulen, Nicolaas M. J.; Mahoney, Walt; Steele, Matt; Domingo, Gonzalo J.

doi:10.1111/j.1365-3156.2011.02773.x

Cited by 18 publications

(14 citation statements)

References 27 publications

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“…These authors also used glycogen/acetate precipitation to improve yields of parasite DNA. Others have shown that measurement of both parasite RNA and DNA by qPCR can improve sensitivity and detection of almost 40% more positive cases [11]. Use of these approaches would improve detection rates with the qPCR methods described here.…”

Section: Discussionmentioning

confidence: 93%

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

Lamikanra

Dobaño

Jiménez

et al. 2012

Malar J

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BackgroundEstimation 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.MethodsA 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 Mcnemar’s test and agreement of positive results quantified as Kappa scores. Parasite prevalences between different age groups were compared by Fisher’s 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 Spearman’s 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.ResultsThese 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).ConclusionsThe 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.

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Section: Discussionmentioning

confidence: 93%

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

Lamikanra

Dobaño

Jiménez

et al. 2012

Malar J

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“…Because of these attributes, circulating levels of Pf HRP-2 reflect the total cumulative parasite biomass [11] and could serve as an indicator of the magnitude of recent (and potentially already cured) infection. Molecular methods are considerably more sensitive than most diagnostic methods, and we have used an approach that has superior sensitivity as a result of detecting the combined RNA and DNA of 18S rRNA gene, allowing us to detect submicroscopic parasitemia as low as 0.02 parasite/µL [5], [21]. Like microscopy and p LDH, but unlike Pf HRP-2, qPCR only detect current parasitemias and therefore cannot account for diurnal fluctuation or for mature trophozoite and schizont stages that are unavailable in the peripheral circulation.…”

Section: Discussionmentioning

confidence: 99%

Comparison of PfHRP-2/pLDH ELISA, qPCR and Microscopy for the Detection of Plasmodium Events and Prediction of Sick Visits during a Malaria Vaccine Study

et al. 2013

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BackgroundCompared to expert malaria microscopy, malaria biomarkers such as Plasmodium falciparum histidine rich protein-2 (PfHRP-2), and PCR provide superior analytical sensitivity and specificity for quantifying malaria parasites infections. This study reports on parasite prevalence, sick visits parasite density and species composition by different diagnostic methods during a phase-I malaria vaccine trial.MethodsBlood samples for microscopy, PfHRP-2 and Plasmodium lactate dehydrogenase (pLDH) ELISAs and real time quantitative PCR (qPCR) were collected during scheduled (n = 298) or sick visits (n = 38) from 30 adults participating in a 112-day vaccine trial. The four methods were used to assess parasite prevalence, as well as parasite density over a 42-day period for patients with clinical episodes.ResultsDuring scheduled visits, qPCR (39.9%, N = 119) and PfHRP-2 ELISA (36.9%, N = 110) detected higher parasite prevalence than pLDH ELISA (16.8%, N = 50) and all methods were more sensitive than microscopy (13.4%, N = 40). All microscopically detected infections contained P. falciparum, as mono-infections (95%) or with P. malariae (5%). By qPCR, 102/119 infections were speciated. P. falciparum predominated either as monoinfections (71.6%), with P. malariae (8.8%), P. ovale (4.9%) or both (3.9%). P. malariae (6.9%) and P. ovale (1.0%) also occurred as co-infections (2.9%). As expected, higher prevalences were detected during sick visits, with prevalences of 65.8% (qPCR), 60.5% (PfHRP-2 ELISA), 21.1% (pLDH ELISA) and 31.6% (microscopy). PfHRP-2 showed biomass build-up that climaxed (1813±3410 ng/mL SD) at clinical episodes.Conclusion PfHRP-2 ELISA and qPCR may be needed for accurately quantifying the malaria parasite burden. In addition, qPCR improves parasite speciation, whilst PfHRP-2 ELISA is a potential predictor for clinical disease caused by P. falciparum.Trial RegistrationClinicalTrials.gov NCT00666380

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“…The use of RDTs for malaria diagnosis increased after the World Health Organization (WHO)’s recommendation to treatment only after a parasitological diagnosis [ 1 ] RDTs are easy to use, do not require electricity or intense expertise and provide results that can be used to make treatment decisions [ 2 ]. However, neither microscopy nor RDTs are able to detect low density parasite infections [ 3 , 4 ] and (reviewed in [ 5 ]) which are often asymptomatic [ 6 – 8 ]. Studies have demonstrated that asymptomatic malaria infections can serve as transmission foci in both low and high transmission settings and, therefore, remain as sources of new infections [ 9 – 13 ].…”

Section: Introductionmentioning

confidence: 99%

PET-PCR method for the molecular detection of malaria parasites in a national malaria surveillance study in Haiti, 2011

Lucchi

Karell

Journel³

et al. 2014

Malar J

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BackgroundRecently, a real-time PCR assay known as photo-induced electron transfer (PET)-PCR which relies on self-quenching primers for the detection of Plasmodium spp. and Plasmodium falciparum was described. PET-PCR assay was found to be robust, and easier to use when compared to currently available real-time PCR methods. The potential of PET-PCR for molecular detection of malaria parasites in a nationwide malaria community survey in Haiti was investigated.MethodsDNA from the dried blood spots was extracted using QIAGEN methodology. All 2,989 samples were screened using the PET-PCR assay in duplicate. Samples with a cycle threshold (CT) of 40 or less were scored as positive. A subset of the total samples (534) was retested using a nested PCR assay for confirmation. In addition, these same samples were also tested using a TaqMan-based real-time PCR assay.ResultsA total of 12 out of the 2,989 samples screened (0.4%) were found to be positive by PET-PCR (mean CT value of 35.7). These same samples were also found to be positive by the nested and TaqMan-based methods. The nested PCR detected an additional positive sample in a subset of 534 samples that was not detected by either PET-PCR or TaqMan-based PCR method.ConclusionWhile the nested PCR was found to be slightly more sensitive than the PET-PCR, it is not ideal for high throughput screening of samples. Given the ease of use and lower cost than the nested PCR, the PET-PCR provides an alternative assay for the rapid screening of a large number of samples in laboratory settings.

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Malaria prevalence defined by microscopy, antigen detection, DNA amplification and total nucleic acid amplification in a malaria‐endemic region during the peak malaria transmission season

Cited by 18 publications

References 27 publications

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

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

Comparison of PfHRP-2/pLDH ELISA, qPCR and Microscopy for the Detection of Plasmodium Events and Prediction of Sick Visits during a Malaria Vaccine Study

PET-PCR method for the molecular detection of malaria parasites in a national malaria surveillance study in Haiti, 2011

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