Identification of Leishmania (Viannia) species and clinical isolates of Leishmania (Leishmania) amazonensis from Brazil using PCR-RFLP of the heat-shock protein 70 gene reveals some unexpected observations

Espada, Caroline R.; Ortíz, Paola A.; Shaw, Jeffrey Jon; Barral, Aldina; Costa, Jackson Maurício Lopes; Uliana, Sílvia Reni Bortolin; Coelho, Adriano C.

doi:10.1016/j.diagmicrobio.2018.03.004

Cited by 30 publications

(20 citation statements)

References 34 publications

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“…The rRNA internal transcribed spacer 1 (ITS-1) region and hsp70 gene are mostly used as suitable target genes, of which the former is applied mainly in the Old World [6, 11, 12, 14, 17, 19, 27, 34, 37–41]. Although the hsp70 gene is one of the most valuable genetic markers for PCR-RFLP-based species identification, intraspecific polymorphism of RFLP patterns and very similar RFLP profiles among species, which affect species identification, have been reported in some Leishmania species [42]. In this study, other nuclear genes, mpi and 6pgd genes, for which encoding enzymes have been used for MLEE, were shown to be alternative useful targets for classification by PCR-RFLP analysis.…”

Section: Discussionmentioning

confidence: 99%

“…Further sample analyses from different areas and different countries will be important to confirm the utility of this analysis, although polymorphic RFLP profiles may be detectable in these genes. Since polymorphism was also reported in the hsp70 gene of several Leishmania species [42], PCR-RFLP analyses of multiple target genes, rather than single nuclear or kinetoplast genes, will result in more accurate species identification and disclose more detailed genetic characteristics of the parasite.…”

Section: Discussionmentioning

confidence: 99%

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PCR-RFLP analyses of Leishmania species causing cutaneous and mucocutaneous leishmaniasis revealed distribution of genetically complex strains with hybrid and mito-nuclear discordance in Ecuador

et al. 2019

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PCR-Restriction Fragment Length Polymorphism (RFLP) analyses targeting multiple nuclear genes were established for the simple and practical identification of Leishmania species without using expensive equipment. This method was applied to 92 clinical samples collected at 33 sites in 14 provinces of Ecuador, which have been identified at the species level by the kinetoplast cytochrome b ( cyt b) gene sequence analysis, and the results obtained by the two analyses were compared. Although most results corresponded between the two analyses, PCR-RFLP analyses revealed distribution of hybrid strains between Leishmania (Viannia) guyanensis and L . (V . ) braziliensis and between L . (V . ) guyanensis and L . (V . ) panamensis , of which the latter was firstly identified in Ecuador. Moreover, unexpected parasite strains having the kinetoplast cyt b gene of L . (V . ) braziliensis and nuclear genes of L . (V . ) guyanensis , L . (V . ) panamensis , or a hybrid between L . (V . ) guyanensis and L . (V . ) panamensis were identified. This is the first report of the distribution of a protozoan parasite having mismatches between kinetoplast and nuclear genes, known as mito-nuclear discordance. The result demonstrated that genetically complex Leishmania strains are present in Ecuador. Since genetic exchanges such as hybrid formation were suggested to cause higher pathogenicity in Leishmania and may be transmitted by more species of sand flies, further country-wide epidemiological studies on clinical symptoms, as well as transmissible vectors, will be necessary.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

PCR-RFLP analyses of Leishmania species causing cutaneous and mucocutaneous leishmaniasis revealed distribution of genetically complex strains with hybrid and mito-nuclear discordance in Ecuador

et al. 2019

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“…Over the last two decades, a multitude of diagnostic assays targeting various genes including the ITS-1, Hsp70, antigen-encoding genes gp63 and cpb, RNA polymerase II, and the N-acetylglucosamine-1phosphate transferase have been designed for the detection and identification of Leishmania species. [41][42][43][44][45] In the absence of a globally accepted "gold standard" for diagnosis, the downside of such an immense range of tests is that many of these methods are obscurely used in very few or individual laboratories throughout the world. Cited more than 185 times in PubMed (viewed March 23, 2019), and used for the detection and identification of various Leishmania species in 63 articles (Supplemental File 2), the ITS-1 remains the most extensively used tool for the diagnosis of leishmaniasis.…”

Section: Discussionmentioning

confidence: 99%

Identification of Clinical Infections of Leishmania Imported into Australia: Revising Speciation with Polymerase Chain Reaction-RFLP of the Kinetoplast Maxicircle

Kaufer

Ellis

Stark

2019

The American Journal of Tropical Medicine and Hygiene

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Leishmaniasis is a vector-borne disease caused by protozoan parasites of the Leishmania genus. In Australia, leishmaniasis is an imported disease that is presenting itself at increased rates because of international travel, the influx of immigrants, and deployment of military operations to endemic regions. Although Leishmania species are morphologically indistinguishable, there is a strong correlation between some causative species of leishmaniasis and the subsequent response to the treatments available and patient outcome. Consequently, identification of the infective species is imperative as misidentification can result in the administering of an ineffective drug. The aim of this study was to develop a simple diagnostic tool with high sensitivity and specificity, which is capable of detecting the presence of the parasite and accurately differentiating the causative species in question. Using the advantageous properties of the maxi-circle kinetoplast DNA, a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) targeting the ND7 gene was developed for the analysis of imported cases of human leishmaniasis in Australia. Designed as a dual analysis, concurrent PCR of Leishmania maxi-circle DNA and digestion with two separate enzymes (NlaIII and HpyCH4IV), this study provides an appraisal on 24 imported cases of leishmaniasis between 2008 and 2017. Five Leishmania species were reported, with members of the Viannia subgenus being the most common. The implementation of novel diagnostic procedures for leishmaniasis such as the one reported here is needed to establish a gold standard practice for the diagnosis and treatment of leishmaniasis.

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“…( V .) lindenbergi ), whilst different strains and isolates of the same species may have different profiles with the same restriction enzyme, leading to misidentification [ 119 ]. Alternatively, Leishmania typing using the hsp70 gene may be solved by DNA sequencing [ 119 ] or HRM [ 57 ] (see below).…”

Section: Molecular Methods For Diagnosis Of Leishmaniasismentioning

confidence: 99%

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

et al. 2020

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Leishmaniasis is a neglected tropical disease with two main clinical forms: cutaneous and visceral leishmaniasis. Diagnosis of leishmaniasis is still a challenge, concerning the detection and correct identification of the species of the parasite, mainly in endemic areas where the absence of appropriate resources is still a problem. Most accessible methods for diagnosis, particularly in these areas, do not include the identification of each one of more than 20 species responsible for the disease. Here, we summarize the main methods used for the detection and identification of leishmaniasis that can be performed by demonstration of the parasite in biological samples from the patient through microscopic examination, by in vitro culture or animal inoculation; by molecular methods through the detection of parasite DNA; or by immunological methods through the detection of parasite antigens that may be present in urine or through the detection of specific antibodies against the parasite. Potential new methods that can be applied for laboratory diagnosis of leishmaniasis are also discussed.

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Identification of Leishmania (Viannia) species and clinical isolates of Leishmania (Leishmania) amazonensis from Brazil using PCR-RFLP of the heat-shock protein 70 gene reveals some unexpected observations

Cited by 30 publications

References 34 publications

PCR-RFLP analyses of Leishmania species causing cutaneous and mucocutaneous leishmaniasis revealed distribution of genetically complex strains with hybrid and mito-nuclear discordance in Ecuador

PCR-RFLP analyses of Leishmania species causing cutaneous and mucocutaneous leishmaniasis revealed distribution of genetically complex strains with hybrid and mito-nuclear discordance in Ecuador

Identification of Clinical Infections of Leishmania Imported into Australia: Revising Speciation with Polymerase Chain Reaction-RFLP of the Kinetoplast Maxicircle

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

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