Structure of
            <b>
              <i>Leishmania</i>
              Minicircle Kinetoplast DNA Classes
            </b>

Yurchenko, Vyacheslav; Merzlyak, Ekaterina M.; Kolesnikov, Alexander; Martinkina, Larissa P.; Vengerov, Yu. Yu.

doi:10.1128/jcm.37.5.1656-1657.1999

Cited by 13 publications

(3 citation statements)

References 9 publications

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“…The high PCR sensitivity in CL is attributed to increased gene copies that are reported to be around 10,000 copies per parasite. 13 Thus, PCR is highly useful in cases where histomorphology only supports a diagnosis of CL as the organism is difficult to demonstrate. We used ITS1 and ITS2 as panfungal primers in confirming and typing cutaneous fungal lesions.…”

Section: Discussionmentioning

confidence: 99%

“…The other primers included:panfungal (ITS1, forward 5′-TCCGTAGGTGAACCTGCGG-3′ and ITS2, reverse 5′-GCTGCGTTCTTCATCGATGC-3′); 11 zygomycetes (ZM1, 5′ATTACCATGAGCAAATCAGA-3′ and ZM3, 5′CAATCC AAGAATTTCACCTCTAG-3′); 12 andleishmaniasis (kDNA, forward GGGKAGGGGCCGTTCTSCGAA and reverse SSSWCTATWTTACACCAACCCC). 13 …”

Section: Methodsmentioning

confidence: 99%

“…panfungal (ITS1, forward 5 0 -TCCGTAGGTGAA CCTGCGG-3 0 and ITS2, reverse 5 0 -GCTGCGTTCTT CATCGATGC-3 0 ); 11 2. zygomycetes (ZM1, 5 0 ATTACCATGAGCAAATCAGA-3 0 and ZM3, 5 0 CAATCC AAGAATTTCACCTCTAG-3 0 ); 12 and 3. leishmaniasis (kDNA, forward GGGKAGGGGCCGTTC TSCGAA and reverse SSSWCTATWTTACACCAA CCCC). 13 The PCR mixture consisted of a total volume of 100 mL containing 8 mL of PCR buffer (•10), 2 mL of 10 mM dNTP blend (2 mM each of dATP, dTTP, dCTP, and dGTP), 2 mL of forward primer (10 pM), 2 mL of reverse primer (10 pM), 1 mL of 3U Taq polymerase, 8 mL of MgCl 2 (25 mM), and 77 mL of Milli-Q water. The PCR cycle included an initial denaturation at 958C for 10 minutes, followed by 3 steps as follows: (1) denaturation phase, 948C for 10 seconds; (2) annealing phase, 638C for 15 seconds; and (3) extension phase, 728C for 25 seconds, with a final extension at 728C for a half-minute and holding it at 208C for 5 minutes.…”

Section: Dna Extraction and Pcrmentioning

confidence: 99%

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Diagnostic Utility of Biplex/Multiplex Polymerase Chain Reaction in Infectious Granulomatous Dermatitis in North Indian Population

Parkhi

S²,

De³

et al. 2020

The American Journal of Dermatopathology

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Background: A definite diagnosis of infectious granulomatous dermatitis (IGD) is difficult for both practicing dermatologists and dermatopathologists due to overlapping clinical and histomorphological features. We aimed to explore the role of multiplex polymerase chain reaction (PCR) for identifying a definite etiological agent for diagnosis and appropriate treatment in IGD in formalinfixed paraffin-embedded tissue.Materials and Methods: Sixty-two cases of IGD were included, excluding leprosy. The histochemical stains including Ziehl-Neelsen, periodic acid-Schiff, and Giemsa were performed in all cases. A multiplex PCR was designed for detection of tuberculosis (TB) (IS6110 and mpt64), fungal infections (ITS1, ITS2; ZM1, and ZM3), and leishmaniasis (kDNA). The results of histomorphology, histochemical stains, and multiplex PCR were compared.Results: Among 62 cases, the sensitivity rate of PCR detection for organisms was 16.7%, 0%, 100%, 72%, 75%, and 66.7% in patients with TB, suggestive of TB, leishmaniasis, fungal infections, and granulomatous dermatitis not otherwise specified and granulomatous dermatitis suggestive of fungus, respectively. The TB PCR using IS6110 primers was negative in all cases; however, PCR using mpt64 primers was positive in 33.33% cases of scrofuloderma. The histochemical stains including Ziehl-Neelsen for acid-fast bacilli, periodic acid-Schiff for fungus, and Giemsa for Leishman-Donovan bodies showed positivity in 11.3%, 43.5%, and 3.2%, respectively. Conclusion:A multiplex PCR (Mycobacterium tuberculosis, Leishmania, and panfungal) is highly recommended in all cases of IGD where an etiological agent is difficult to establish by skin biopsy and histochemical stains along with a clinicopathological correlation. This will augment in appropriate treatment and will reduce empirical treatment and morbidity in such patients.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Dna Extraction and Pcrmentioning

confidence: 99%

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Diagnostic Utility of Biplex/Multiplex Polymerase Chain Reaction in Infectious Granulomatous Dermatitis in North Indian Population

Parkhi

S²,

De³

et al. 2020

The American Journal of Dermatopathology

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Homologous minicircles in Leishmania donovani

Lambson

Barker

2002

Transactions of the Royal Society of Tropical Medicine and Hygi

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Leishmania minicircular deoxyribonucleic acid (DNA) is arranged into different classes according to sequence. These classes differ substantially in sequence, despite species- and genus-specific regions, and are present in widely different copy numbers within and between Leishmania strains. Homologous minicircles have been identified in different species of Leishmania by comparing sequences of known minicircles. However, it is possible to select for minicircles of the same class by amplifying Leishmania DNA with polymerase chain reaction primers from the conserved and variable regions. This approach was used with 2 different minicircle classes in the L. donovani complex. In all isolates tested it was possible to amplify minicircles of the selected class.

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Complete minicircle genome ofLeptomonas pyrrhocorisreveals sources of its non-canonical mitochondrial RNA editing events

Gerasimov

Gasparyan

Afonin

et al. 2021

Nucleic Acids Research

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Uridine insertion/deletion (U-indel) editing of mitochondrial mRNA, unique to the protistan class Kinetoplastea, generates canonical as well as potentially non-productive editing events. While the molecular machinery and the role of the guide (g) RNAs that provide required information for U-indel editing are well understood, little is known about the forces underlying its apparently error-prone nature. Analysis of a gRNA:mRNA pair allows the dissection of editing events in a given position of a given mitochondrial transcript. A complete gRNA dataset, paired with a fully characterized mRNA population that includes non-canonically edited transcripts, would allow such an analysis to be performed globally across the mitochondrial transcriptome. To achieve this, we have assembled 67 minicircles of the insect parasite Leptomonas pyrrhocoris, with each minicircle typically encoding one gRNA located in one of two similar-sized units of different origin. From this relatively narrow set of annotated gRNAs, we have dissected all identified mitochondrial editing events in L. pyrrhocoris, the strains of which dramatically differ in the abundance of individual minicircle classes. Our results support a model in which a multitude of editing events are driven by a limited set of gRNAs, with individual gRNAs possessing an inherent ability to guide canonical and non-canonical editing.

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Structure of Leishmania Minicircle Kinetoplast DNA Classes

Cited by 13 publications

References 9 publications

Diagnostic Utility of Biplex/Multiplex Polymerase Chain Reaction in Infectious Granulomatous Dermatitis in North Indian Population

Diagnostic Utility of Biplex/Multiplex Polymerase Chain Reaction in Infectious Granulomatous Dermatitis in North Indian Population

Homologous minicircles in Leishmania donovani

Complete minicircle genome ofLeptomonas pyrrhocorisreveals sources of its non-canonical mitochondrial RNA editing events

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