Malaria in pregnant women living in areas of low transmission on the southeast Brazilian Coast: molecular diagnosis and humoural immunity profile

Abstract

Studies on autochthonous malaria in low-transmission areas in Brazil have acquired epidemiological relevance because they suggest continued transmission in what remains of the Atlantic Forest. In the southeastern portion of the state of São Paulo, outbreaks in the municipality of Juquitiba have been the focus of studies on the prevalence of Plasmodium, including asymptomatic cases. Data on the occurrence of the disease or the presence of antiplasmodial antibodies in pregnant women from this region have not previously been described. Although Plasmodium falciparum in pregnant women has been widely addressed in the literature, the interaction of Plasmodium vivax and Plasmodium malariae with this cohort has been poorly explored to date. We monitored the circulation of Plasmodium in pregnant women in health facilities located in Juquitiba using thick blood film and molecular protocols, as well as immunological assays, to evaluate humoural immune parameters. Through real-time and nested polymerase chain reaction, P. vivax and P. malariae were detected for the first time in pregnant women, with a positivity of 5.6%. Immunoassays revealed the presence of IgG antibodies: 44% for ELISA-Pv, 38.4% for SD-Bioline-Pv and 18.4% for indirect immunofluorescence assay-Pm. The high prevalence of antibodies showed significant exposure of this population to Plasmodium. In regions with similar profiles, testing for a malaria diagnosis might be indicated in prenatal care.

Serological and molecular techniques applied for identification of Plasmodium spp. in blood samples from nonhuman primates

“…In all the reactions, ultra-pure water (Invitrogen  , Carlsbad, CA, USA) was used as a negative control. Plasmodium vivax and P. malariae DNA samples from human infected patients (HRISTOV et al, 2014;LIMA et al, 2016) and P. falciparum DNA (strains K1 and Palo Alto obtained through in vitro culturing) were used as positive controls. The products were then subjected to a second reaction, using 2 μL of the amplified product from the first reaction, in a final reaction volume of 25 μL, containing 1X Taq buffer, 2.5 mM of MgCl 2 , 0.1 mM of deoxynucleotide triphosphate mixture, 0.5 μM of PLF, 0.064 μM of MAR, 0.3 μM of FAR, 0.05 μM of VIR and 1.25 U of Taq DNA polymerase.…”

“…Construction of plasmid standards containing Plasmodium sp. 18S rRNA gene fragments Plasmodium falciparum and P. malariae (HRISTOV et al, 2014;LIMA et al, 2016) 18S rRNA gene fragments were amplified in conventional PCR assays using the same set of primers that are used in qPCR assays (GAMA et al, 2007). The amplification reactions were performed in a T100 thermal cycler (Bio Rad), using 4 μL of DNA in a 46 μL mixture, containing 1X PCR buffer, 2 mM of MgCl 2 , 1.25 U of Taq Platinum DNA Polymerase (Life Technologies  , Carlsbad, CA, USA), 200 μM of each dNTP and 200 nM of each M60 and M61 primer.…”

“…IFAT was performed using the antigen from crude human erythrocyte infected with schizonts of P. malariae (HRISTOV et al, 2014) and P. falciparum (K1 and Palo Alto strains), obtained from in vitro cultures. Plasmodium berghei ANKA antigen was obtained from red cells from BALB/c experimentally infected mice.…”

Polimorfismos genéticos em genes relacionados com imunidade inata em população de área de baixa endemicidade para malária

“…Esses casos ocorrem com baixos níveis de parasitemia causados por P. vivax [191][192][193][194][195][196] e P. malariae [197][198][199][200] Controle Caso Controle Caso Controle Caso HH=0 47 74 24 67 15 38 Hh, hh=1 16 58 39 65 48 94 A idade foi centralizada pela média (38 anos) e o sexo foi codificado em "0" para as mulheres e "1" para os homens.…”

Alterações histopatológicas placentárias associadas à infecção por <i>Plasmodium vivax</i> em gestantes do Vale do Alto Juruá.

“…Embora existam trabalhos realizados com gestantes da região Amazônica brasileira (CARVALHO et al, 2011;CHAGAS et al, 2009;JARUDE;TRINDADE;TAVARES-NETO, 2003;MACHADO FILHO et al, 2014; MARTÍNEZ-ESPINOSA; DANIEL-RIBEIRO; ALECRIM, 2004) e da região extra-Amazônica (HRISTOV et al, 2014), poucos foram conduzidos com objetivo de avaliar a fisiopatologia da malária gestacional e placentária no Brasil (ATAÍDE et al, 2015;CARVALHO et al, 2010;SOUZA et al, 2013).…”