Natural Infection of<i>Nyssomyia neivai</i>by<i>Leishmania (Viannia</i>) spp. in the State of Paraná, Southern Brazil, Detected by Multiplex Polymerase Chain Reaction

Oliveira, Diego Molina de; Reinhold-Castro, Kárin Rosi; Bernal, Marcos Vinícius Zandonadi; Legriffon, Cristiane Machado de Oliveira; Lonardoni, Maria Valdrinez Campana; Teodoro, Ueslei; Silveira, Thaís Gomes Verzignassi

doi:10.1089/vbz.2009.0218

Cited by 23 publications

(23 citation statements)

References 40 publications

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“…intermedia suggest that it also is associated with Leishmania transmission in this region. Both species have been found to be naturally infected with Leishmania and are associated with ATL foci (Forattini et al 1972, Rangel et al 1984, Casanova et al 1995, Córdoba-Lanus et al 2006, Marcondes et al 2009, Saraiva et al 2009, Pita-Pereira et al 2009, Margonari et al 2010, Oliveira et al 2011, thereby demonstrating the great epidemiological importance of these species. Lutz and Neiva (1912) have reported that Ny.…”

Section: Discussionmentioning

confidence: 99%

Biogeographical aspects of the occurrence of Nyssomyia neivai and Nyssomyia intermedia (Diptera: Psychodidae) in a sympatric area of the Brazilian savannah

Saraiva¹,

Carvalho²,

Sanguinette³

et al. 2012

Mem. Inst. Oswaldo Cruz

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Section: Discussionmentioning

confidence: 99%

Biogeographical aspects of the occurrence of Nyssomyia neivai and Nyssomyia intermedia (Diptera: Psychodidae) in a sympatric area of the Brazilian savannah

Saraiva¹,

Carvalho²,

Sanguinette³

et al. 2012

Mem. Inst. Oswaldo Cruz

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“…Extraction of DNA: The samples were macerated, and DNA was extracted with a solution of guanidine isothiocyanate and phenol, and hydrated in 20 µL of ultra-pure H 2 O 31 . For each group of 22 samples submitted for DNA extraction, one negative control (male sandflies) and one positive control [male sandflies plus 10 4 L. (V.) braziliensis promastigotes] were used.…”

Section: Methodsmentioning

confidence: 99%

“…The amplification was carried out in a PC Thermocycler (Biometra, Germany) at 94 °C for seven min to activate the enzyme, followed by 30 cycles, each divided into three stages, of denaturation (1.5 min at 95 °C), annealing (1.5 min at 57 °C), and polymerization (two min at 72 °C). After this, the extension was continued for a further 10 min at 72 °C, and the tubes were then kept at 4 °C until analysis 31 . The amplification products were submitted to electrophoresis in 2% of agarose gel (Invitrogen, Paisley, Scotland, UK) stained with 0.1 µg/mL ethidium bromide, at 10-15 V/cm.…”

Section: Methodsmentioning

confidence: 99%

DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL

Neitzke-Abreu

Reinhold-Castro

Venazzi

et al. 2014

Rev. Inst. Med. trop. S. Paulo

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Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.

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“…such as the quantitative polymerase chain reaction (qPCR), nested PCR, and multiplex PCR (20)(21)(22)(23)(24)(25)(26)(27)(28). The possibility of combining multiple targets in the same assay led to expansion of applications of the multiplex PCR technique on leishmaniasis diagnosis, since the identification of complexes and species of the parasite to evaluation of sample integrity and PCR performance on sandfly pools (23)(24)(25)(26)(27)(28).…”

Section: Introductionmentioning

confidence: 99%

Application of the mammalian glyceraldehyde-3-phosphate dehydrogenase gene for sample quality control in multiplex PCR for diagnosis of leishmaniasis

Gonçalves¹,

Régis-da-Silva²,

Brito³

et al. 2012

J. Venom. Anim. Toxins incl. Trop. Dis

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Leishmaniasis is a neglected disease endemic in five continents. It is a severe disease that may lead to death, and its early detection is important to avoid severe damage to affected individuals. Molecular methods to detect Leishmania are considered alternatives to overcome the limitations presented by conventional methods. The aim of this study was to develop multiplex PCR systems able to detect small amounts of target DNA of Leishmania infantum and Leishmania braziliensis, and the gene coding for glyceraldehyde-3-phosphate dehydrogenase (G3PD) in mammals, enabling quality evaluation of the sample simultaneously with detection of the specific target. The systems created for G3PD recognition were combined with detection systems for L. infantum and L. braziliensis to compose multiplex PCR systems for visceral (mVL) and cutaneous (mACL) leishmaniasis diagnosis. The multiplex PCR systems developed were assessed in blood samples from five different species of mammal reservoirs involved in the disease cycle in Brazil, and 96 and 52 human samples from patients with suspected visceral leishmaniasis (VL) and cutaneous leishmaniasis (ACL), respectively. Three G3PD detection systems were created (G3PD1, G3PD2 and G3PD3) with different product sizes, G3PD2 was chosen for the formation of multiplex PCR systems. The two multiplex PCR systems (mVL and mACL) were reproducible in all species evaluated. Results of test samples (sensitivity, specificity and efficiency) suggest its use in routine diagnosis, research activities in medicine and veterinary medicine. Additionally, the systems designed to detect the G3PD gene are capable of combining with other targets used for molecular diagnosis of infectious diseases. Concerning leishmaniasis, the multiplex PCR systems can be used in epidemiological studies for the detection of new and classic reservoirs, which may contribute to the reliability of results and development of actions to control the disease.

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Natural Infection ofNyssomyia neivaibyLeishmania (Viannia) spp. in the State of Paraná, Southern Brazil, Detected by Multiplex Polymerase Chain Reaction

Cited by 23 publications

References 40 publications

Biogeographical aspects of the occurrence of Nyssomyia neivai and Nyssomyia intermedia (Diptera: Psychodidae) in a sympatric area of the Brazilian savannah

Biogeographical aspects of the occurrence of Nyssomyia neivai and Nyssomyia intermedia (Diptera: Psychodidae) in a sympatric area of the Brazilian savannah

DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL

Application of the mammalian glyceraldehyde-3-phosphate dehydrogenase gene for sample quality control in multiplex PCR for diagnosis of leishmaniasis

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