Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale

Lima, Giselle F M C; Levi, José Eduardo; Geraldi, Marcelo Plaisant; Sanchez, Maria Carmen Arroyo; Segurado, Aluísio Cotrim; Hristov, Angelica D.; Inoue, Juliana; Costa-Nascimento, Maria de Jesus; Santi, Sílvia Maria Di

doi:10.1590/s0074-02762011000600008

Cited by 42 publications

(67 citation statements)

References 34 publications

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“…When RDTs were used as the reference standard, nPCR exhibited sensitivity and positive values of 100% and 39.38% respectively. This ability for PCR to more accurately detect low-grade parasitemia with a higher degree of sensitivity than microscopy and RDTs supports the reports by other researchers [22], [27][28][29][30]. The high false negative obtained using RDTs has been reported and suggested that all RDT negative results be confirmed by microscopic analysis [31].…”

Section: Discussionsupporting

confidence: 86%

Comparison of Nested PCR and Conventional Analysis of <i>Plasmodium</i> Parasites in Kano, Nigeria

Vincent¹

2017

EJCBS

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Plasmodium identification represents the crucial factor in malaria diagnosis and treatment across developing countries. Conventional microscopy and the use of rapid diagnostic kits have been extensively applied towards human malaria diagnosis. Recombinant DNA techniques have been applied towards malaria diagnosis as well as in the species specific identification using Plasmodium 18s-rRNA gene. This study was undertaken amongst patients attending the Murtala Mohammed Specialist Hospital, Kano. Blood samples were collected from 350 malaria suspected patients. Microscopic analysis via Giemsa-staining revealed that 220 patients were positive for malaria. RDT analysis showed that 248 test samples were positive for Plasmodium infection. DNA products obtained from the blood samples were analyzed by nested PCR to amplify the 18S ssrRNA Plasmodium gene with genus and specific primers rPLU1/5, rPLU3/4, rVIV1/2, rFAL1/2, rMAL1/2 and rOVA1/2. Data obtained showed that 58.64% of specimens tested by microscopy were false positives while 60.62% of false positives were obtained using RDTs in comparison to nPCR which proved that on 91 out of 350 patients were infected with Plasmodium falciparum, representing 26% of tested specimen. Comparative analysis of nPCR to microscopy showed that the sensitivity and positive predictive values of the nPCR were determined as 100 and 41.36%, respectively, while against RDTs it was 100 and 39.38% respectively. nPCR was determined to be more sensitive and specific than either microscopy or RDTs. This study revealed that the accurate diagnosis of malaria by nPCR was compulsory in malaria-prone regions of Nigeria such that nPCR should be applied routinely in laboratory studies.

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Section: Discussionsupporting

confidence: 86%

Comparison of Nested PCR and Conventional Analysis of <i>Plasmodium</i> Parasites in Kano, Nigeria

Vincent¹

2017

EJCBS

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“…One sample was positive for P. malariae by PCR [19]. Recently, samples of 39 donors from the same area were assayed employing a modified real-time PCR protocol [20] using genus-specific primers [17]. ELISA- Pv was reagent in 38% of samples and SD Bioline Pf / Pv was positive in 38% ( Pv band) and 0% ( Pf band).…”

Section: Introductionmentioning

confidence: 99%

Unexpected detection of Plasmodium vivax and Plasmodium falciparum DNA in asymptomatic blood donors: fact or artifact?

Mendrone

Cerutti

Levi

et al. 2014

Malar J

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A study searching for Plasmodium vivax and Plasmodium falciparum DNA among blood donors from the non-endemic area in Brazil reported a rate of 7.41%. This number is at least three times higher than what has been observed in blood donors from the Amazon, an endemic area concentrating >99% of all malaria cases in Brazil. Moreover, the majority of the donors were supposedly infected by P. falciparum, a rare finding both in men and anophelines from the Atlantic forest. These findings shall be taken with caution since they disagree with several publications in the literature and possibly overestimate the actual risk of malaria transmission by blood transfusion in São Paulo city.

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“…PCR techniques are highly sensitive for the detection of parasite nucleic acids [7][8][9], are particularly valuable for species confirmation and should be used in all instances when species cannot be determined by microscopy or to evaluate for mixed infections [5] but unfortunately are not always available everywhere. Immunochromatographic tests most often using a dipstick or a cassette format contain monoclonal antibodies against parasite specific antigens and provide results in 5-20 minutes [10,11].…”

Section: Discussionmentioning

confidence: 99%

Difficulties in Diagnosis of Malaria in Non-Endemic Areas: A Case Report of a Child in Brazil

Brustoloni¹,

Baldissera²,

Brustoloni³

2015

Malaria Contr Elimination

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Malaria is a tropical parasitic disease that triggers severe social and economic problems worldwide and causes thousands of deaths each year. Early diagnosis enables proper treatment. People from non-endemic areas who travel to areas of transmission are often exposed to late or incorrect diagnosis because healthcare professionals have great difficulty in conceiving malaria as a diagnostic hypothesis. In addition, technicians skilled in recognizing malaria etiologic agent are not abundant. This paper addresses the case of a child from a non-endemic malaria region in Brazil, with a prior hypothetic diagnosis of visceral leishmaniasis, however bone marrow aspirate revealed Plasmodium instead of Leishmania. A retrospective epidemiological investigation showed a history of a recent trip to a malaria area and the review of blood smears obtained on admission disclosed a large number of trophozoites and gametocytes of Plasmodium falciparum that had been unnoticed, suggesting that diagnosis could have been possible earlier. Malaria clinical picture is easily mistaken for other febrile infectious diseases, thus the possibility of its occurrence cannot be ruled out. There is a need to increasingly train the microscopy technicians, both in endemic and non-endemic areas, so as to minimize the false diagnoses and decrease morbidity and mortality of the disease.

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Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale

Cited by 42 publications

References 34 publications

Comparison of Nested PCR and Conventional Analysis of <i>Plasmodium</i> Parasites in Kano, Nigeria

Comparison of Nested PCR and Conventional Analysis of <i>Plasmodium</i> Parasites in Kano, Nigeria

Unexpected detection of Plasmodium vivax and Plasmodium falciparum DNA in asymptomatic blood donors: fact or artifact?

Difficulties in Diagnosis of Malaria in Non-Endemic Areas: A Case Report of a Child in Brazil

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Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale

Cited by 42 publications

References 34 publications

Comparison of Nested PCR and Conventional Analysis of &lt;i&gt;Plasmodium&lt;/i&gt; Parasites in Kano, Nigeria

Comparison of Nested PCR and Conventional Analysis of &lt;i&gt;Plasmodium&lt;/i&gt; Parasites in Kano, Nigeria

Unexpected detection of Plasmodium vivax and Plasmodium falciparum DNA in asymptomatic blood donors: fact or artifact?

Difficulties in Diagnosis of Malaria in Non-Endemic Areas: A Case Report of a Child in Brazil

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Comparison of Nested PCR and Conventional Analysis of <i>Plasmodium</i> Parasites in Kano, Nigeria

Comparison of Nested PCR and Conventional Analysis of <i>Plasmodium</i> Parasites in Kano, Nigeria