Rapid and sensitive detection of <i>Leishmania</i> kinetoplast DNA from spleen and blood samples of kala-azar patients

Smyth, Audra J.; Ghosh, Anil K.; Hassan, Md. Quamarul; Basu, Debasis; Bruijn, M.H.L. de; Adhya, Samit; Mallik, K. K.; Barker, Douglas C.

doi:10.1017/s0031182000074096

Cited by 176 publications

(101 citation statements)

References 13 publications

(9 reference statements)

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“…However, despite using primers for kDNA targets, Smyth et al (1992) found the same sensitivity (1 pg/µL) reported in the present study. Although most authors have used the kinetoplast minicircle region for primer construction, the present study showed that primers selected from 18S rRNA are a reliable option for specific molecular detection of L. infantum chagasi.…”

Section: Resultssupporting

confidence: 83%

Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

Neto

Duarte

Costa

et al. 2012

Rev. Bras. Parasitol. Vet.

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The objective of this study was to design and evaluate new primers for species-specific detection of L. infantum chagasi using PCR. Two combinations of primer pairs were established with the aim of obtaining specific amplification products from the L. infantum chagasi 18S rRNA gene. The combinations of the primer pairs and the respective sizes of the PCR products, based on the U422465 GenBank reference sequence of L. infantum chagasi, were: LCS1/LCS3 (259 bp) and LCS2/LCS3 (820 bp). It was concluded that the new PCR assays optimized using the primer pairs LCS1/LCS3 and LCS2/LCS3 were effective for specific detection of L. infantum chagasi, with analytical sensitivity to detect 1 pg/µL of DNA.

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Section: Resultssupporting

confidence: 83%

Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

Neto

Duarte

Costa

et al. 2012

Rev. Bras. Parasitol. Vet.

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“…Diagnostic methods based on deoxyribonucleic acid (DNA) have, however, now made the sensitive and rapid detection and identification of many microorganisms possible. Over the last couple of decades, for example, assays based on PCR have proved to be sensitive and powerful tools for the detection and identification of Leishmania in clinical samples, without the need to culture the parasites (Rodgers et al, 1990;Smyth et al, 1992;Lopez et al, 1993;Noyes et al, 1998). In the present study, based in Ahvaz (the capital city of Khouzestan), a nested PCR was used to confirm the microscopical diagnosis of CL and identify the causative parasites to species level.…”

mentioning

confidence: 99%

The PCR-based detection and identification of the parasites causing human cutaneous leishmaniasis in the Iranian city of Ahvaz

Ghasemian

Maraghi

Samarbafzadeh

et al. 2011

Annals of Tropical Medicine & Parasitology

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In Iran, Leishmania major or L. tropica cause almost all of the human cutaneous leishmaniasis (CL). Unfortunately, the detection methods frequently used for CL (the microscopical examination of direct smears or the culture of biopsies) are not very sensitive and the Leishmania species causing each case of CL in Iran is usually only tentatively identified from extrinsic factors, such as the case's clinical manifestations and region of residence. Recently, however, a nested PCR that targets the parasites' kinetoplast DNA has been used in the city of Ahvaz (the capital of the province of Khouzestan, in south-western Iran) to confirm the microscopical diagnosis of CL and to identify the causative parasites, to species level. Smears from the lesions on 100 suspected cases of CL were fixed, stained with Wright's eosin-methylene blue, and checked for amastigotes under a light microscope. Scrapings from the same smears were then tested for leishmanial DNA, using a nested PCR that allows the DNA from L. tropica to be identified and distinguished from that of L. major. The 100 smears investigated were all found amastigote-positive by microscopy and PCR-positive for either L. major DNA (97 smears) or L. tropica DNA (three smears). The predominant species causing CL in Ahvaz is therefore L. major.

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“…Primers specific for L. Viannia kDNA (B1/B2) developed by de Brujin and Barker (1992) and for L. chagasi kDNA (DB8/AJS3) developed by Smyth (1992) were used in these assays. PCR products were hybridized with biotinilated probes.…”

Section: Methodsmentioning

confidence: 99%

Susceptibility of spiny rats (Proechimys semispinosus) to Leishmania (Viannia) panamensis and Leishmania (Leishmania) chagasi

Travi

Arteaga

León

et al. 2002

Mem. Inst. Oswaldo Cruz

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The role of Proechimys semispinosus as reservoir of Leishmania (Viannia) Key words: spiny rats -Leishmania -reservoir -experimental infection Spiny rats (Proechimys spp.) are parasitized by a variety of Leishmania species, including L. aristidesi and recognized pathogenic species such as L. guyanensis, L. amazonensis, and L. mexicana s.l. Although these rodents have been incriminated as natural hosts for pathogenic species in several endemic foci in South America, encompassing Brazil, French Guiana, Venezuela and Trinidad (Lainson et al. 1979, WHO 1990, their reservoir role still needs to be established.In Colombia, information on spiny rats and their relationships with both cutaneous and visceral leishmaniasis is fragmentary. P. semispinosus is the most abundant small mammal inhabiting tropical wet forests of the Colombian Pacific coast where cutaneous leishmaniasis is endemic (Weigle et al. 1986, Gonzalez & Alberico 1993. In the Northern region of the country, another species of spiny rat, P. canicollis, is infected with L. chagasi in tropical dry forests and areas of subsistence agriculture where the original forest has been extensively degraded .Incriminating wild animals as Leishmania reservoirs is based on several biological parameters, among which natural infection, population dynamics, and interactions with and infectivity to vectors are of capital importance. In the present study, using an experimental approach, we evaluated the potential role of P. semispinosus as a reservoir of cutaneous leishmaniasis caused by L. (V.) panamensis. Also, we determined the susceptibility of this species to L. (L.) chagasi to evaluate the utility of this rodent as a model of reservoir host that could be utilized in the laboratory. MATERIALS AND METHODSCollection of spiny rats and quarantine -Spiny rats from the Pacific lowlands of Colombia (Tumaco, Nariño; 1°48'N,78°46'W) were captured with National-type traps, after the appropriate permission from the departmental agency for the protection of wild fauna (Corponariño) was obtained. Approximately 100 trapping stations were established in areas of secondary forest, where banana, plantain and cocoa are grown. The traps were baited with plantain, set before dusk, and inspected at dawn. To avoid excessive stress and self-inflicted wounds, the animals were transferred immediately to transport cages of aluminum walls, where access to food and water was provided. Specimens were transported by air to the central laboratory in Cali and quarantined for 8 weeks in the vivarium. An individual coprological analysis was conducted, and an anti-parasitic treatment [ivermectin (Virbamec®), and propoxur (Bolfo®)] was implemented. A blood sample (0.5 ml), drawn by intracardial puncture, was cultured in NNN and Senekjie's media to detect natural infections with Leishmania or Trypanosoma. Serum was collected for detecting antibodies against the same parasites by means of ELISA. Total IgG antibodies were determined in serum (diluted 1:100) using a soluble T. cruzi or L. panamensis

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Rapid and sensitive detection of Leishmania kinetoplast DNA from spleen and blood samples of kala-azar patients

Cited by 176 publications

References 13 publications

Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

The PCR-based detection and identification of the parasites causing human cutaneous leishmaniasis in the Iranian city of Ahvaz

Susceptibility of spiny rats (Proechimys semispinosus) to Leishmania (Viannia) panamensis and Leishmania (Leishmania) chagasi

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