Duplex qPCR for Leishmania species identification using lesion imprint on filter paper.

Abstract:

BACKGROUND:American cutaneous leishmaniasis (ACL) is caused by different Leishmania parasites, which stimulate and direct the immune response against the infection. OBJECTIVE:To evaluate the TaqMan probe technology applicability to diagnose and identifying of Leishmania spp. related to the ACL etiology. METHODOLOGY:Through the MEGA 6.0 software, performed an in silico analysis using multiple alignments of Leishmania spp. which were available on GenBank for different genomic targets. The efficiency (e), specificity and detection limit (DL) were calculated for each system, these were associated to compose a duplex-qPCR (DqPCR). The samples of blood, lesion biopsy and lesion imprint on filter paper from patients residing in states of Amazonas (AM) and Pernambuco (PE)-Brazil, (cases and controls) were used to perform the DqPCR technique. The capacity to identify the Leishmania species was determined by comparison with isoenzymes method and sequencing analysis. RESULTS:Internal Transcribed Spacer 1 (rDNA) was the target selected. Two sets of primers and probes were designed and combined: SVS for subgenus Viannia and LaS for L. (L.) amazonensis. The results were: SVSe = 93.24%, SVS DL = 50 fg/μL; LaSe = 89.3%, LaSLD = 5 fg/μL presented 100% of specificity. In total, 236 individuals participated of the present study, wherein were 101 blood samples, 33 biopsies and 147 lesion imprints. The imprint was the most sensitive sample, showing 83.06% of sensitivity, 86.96% of specificity and substantial agreement between the techniques analysis (k = 0.531; p < 0,001). Regarding the species identification, DqPCR and sequencing/isoenzymes have agreed at 100%, since the infection is caused by a single Leishmania species. CONCLUSION:The DqPCR technique was applicable in diagnosis and identification of Leishmania spp. (subgenus Viannia and L. amazonensis). Furthermore, the lesion imprint is less invasive, allowing a fewer discomfort and greater acceptance by the patients, in addition of being low cost and easy handling.

journal_name

Exp Parasitol

authors

Morais RCS,Melo MGN,Goes TC,Pessoa E Silva R,Morais RF,Guerra JAO,Brito MEF,Brandão-Filho SP,Paiva Cavalcanti M

doi

10.1016/j.exppara.2020.108019

subject

Has Abstract

pub_date

2020-12-01 00:00:00

pages

108019

eissn

0014-4894

issn

1090-2449

pii

S0014-4894(20)30006-0

journal_volume

219

pub_type

杂志文章
  • In vitro growth, cytopathic effects and clearance of monolayers by clinical isolates of Balamuthia mandrillaris in human skin cell cultures.

    abstract::Balamuthia mandrillaris is a free-living ameba (FLA) that has been isolated or its DNA identified in soil, dust and water. It causes a fatal central nervous system infection in humans and animals. Although it is environmental as Acanthamoeba and Naegleria fowleri, the two other free-living amebae that also cause CNS i...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2015.05.004

    authors: Yera H,Dupouy-Camet J,Jackson JW,Sriram R,Sweat S,Goldstein JM,Visvesvara GS

    更新日期:2015-09-01 00:00:00

  • Toxoplasma gondii: in vitro and in vivo activities of the hydroxynaphthoquinone 2-hydroxy-3-(1'-propen-3-phenyl)-1,4-naphthoquinone alone or combined with sulfadiazine.

    abstract::The compound 2-hydroxy-3-(1'-propen-3-phenyl)-1,4-naphthoquinone (PHNQ6) was evaluated for activity against Toxoplasma gondii, alone or combined with sulfadiazine. Treatment with PHNQ6 combined with sulfadiazine protected at least 70 and 90% of mice infected with RH and EGS strains, respectively. Mice were treated wit...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2005.12.006

    authors: Ferreira RA,Oliveira AB,Ribeiro MF,Tafuri WL,Vitor RW

    更新日期:2006-06-01 00:00:00

  • Opisthorchis viverrini: Gene expression profiling of carcinogenic adult liver fluke worms using 5' SAGE.

    abstract::Opisthorchis viverrini is the only liver fluke that has been proved to be associated with cholangiocarcinoma (CCA). However, the mechanisms by which O. viverrini participates in the carcinogenesis of CCA are still unclear. To understand the biology and host-parasite interaction related to O. viverrini infection, gene ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2008.08.004

    authors: Chutiwitoonchai N,Shen Y,Zheng H,Xiong H,Zhao G,Imtawil K,Intapan PM,Wongkham S,Wongkham C

    更新日期:2008-12-01 00:00:00

  • Echinostoma paraensei and Schistosoma mansoni: adherence of unaltered or modified latex beads to hemocytes of the host snail Biomphalaria glabrata.

    abstract::Hemocytes derived from a strain (13-16-R1) of Biomphalaria glabrata resistant to Schistosoma mansoni were significantly more likely to bind untreated latex beads than hemocytes from the schistosome-susceptible M line strain. Beads preincubated in 13-16-R1 plasma were more readily bound by both 13-16-R1 and M line hemo...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(92)90182-a

    authors: Uchikawa R,Loker ES

    更新日期:1992-09-01 00:00:00

  • Novel mutations in the antifolate drug resistance marker genes among Plasmodium vivax isolates exhibiting severe manifestations.

    abstract::Plasmodium vivax is the predominant species of the human malaria parasite present in the Indian subcontinent. There have been recent reports on Chloroquine (CQ) resistance and severe manifestations shown by P. vivax from different regions of the world including India. This study focuses on Bikaner, India where during ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2012.09.018

    authors: Garg S,Saxena V,Lumb V,Pakalapati D,Boopathi PA,Subudhi AK,Chowdhury S,Kochar SK,Kochar DK,Sharma YD,Das A

    更新日期:2012-12-01 00:00:00

  • A high prevalence of Theileria penicillata in woylies (Bettongia penicillata).

    abstract::The woylie or brush-tailed bettong (Bettongia penicillata) is a medium-sized native Australian marsupial that has undergone a dramatic decline in numbers in recent years. Trypanosome parasites have been identified in the woylie but little is known about the prevalence and clinical impact of other haemoprotozoan parasi...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2012.03.013

    authors: Rong J,Bunce M,Wayne A,Pacioni C,Ryan U,Irwin P

    更新日期:2012-06-01 00:00:00

  • Entamoeba invadens: sphingolipids metabolic regulation is the main component of a PKC signaling pathway in controlling cell growth and proliferation.

    abstract::The sphingolipids biosynthesis pathway generates bioactive molecules crucial to the regulation of physiological processes. We have recently reported that DAG (diacylglycerol) generated during sphingomyelin synthesis, plays an important role in PKC (protein kinase C) activation, necessary for the transit through the ce...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2009.02.011

    authors: Cerbon J,Olguin T,Alvarez-Grave PR,López-Sánchez RC

    更新日期:2009-06-01 00:00:00

  • IFN-γ-induced macrophage antileishmanial mechanisms in mice: A role for immunity-related GTPases, Irgm1 and Irgm3, in Leishmania donovani infection in the liver.

    abstract::In C57BL/6 mice, Leishmania donovani infection in the liver provoked IFN-γ-induced expression of the immunity-related GTPases (IRG), Irgm1 and Irgm3. To gauge the antileishmanial effects of these macrophage factors in the liver, intracellular infection was analyzed in IRG-deficient mice. In early- (but not late-) stag...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2015.07.005

    authors: Murray HW,Mitchell-Flack M,Taylor GA,Ma X

    更新日期:2015-10-01 00:00:00

  • Entamoeba histolytica: an ecto-phosphatase activity regulated by oxidation-reduction reactions.

    abstract::In this work, an ecto-phosphatase activity of Entamoeba histolytica was characterized using intact cells. This activity presented the following biochemical characteristics: (i) it hydrolyzes p-NPP with V(max) of 8.00+/-0.22 nmol p-NP x h(-1) x 10(-5) cells and K(m) of 2.68+/-0.25 mM; (ii) it is inhibited by acid phosp...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2006.09.019

    authors: de Sá Pinheiro AA,Amazonas JN,de Souza Barros F,De Menezes LF,Batista EJ,Silva EF,De Souza W,Meyer-Fernandes JR

    更新日期:2007-04-01 00:00:00

  • Peptidases of pinworms Syphacia muris and Passalurus ambiguus.

    abstract::In this first report about pinworms peptidases we primarily characterize peptidases released during in vitro maintenance of two common pinworms of laboratory animals -Syphacia muris and Passalurus ambiguus. The peptidase activity obtained using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showe...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2010.04.018

    authors: Vadlejch J,Lytvynets A,Jankovská I,Langrová I

    更新日期:2010-10-01 00:00:00

  • Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase.

    abstract::Trypanosomatid protozoa lack consensus promoters for RNA polymerase (RNAP) II. However, the artificial insertion of the T7 promoter (P(T7)) and the tetracycline repressor into Trypanosoma brucei cell lines expressing T7RNAP allows P(T7)-driven gene expression to be tetracycline-inducible. These cell lines provide a mo...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2007.01.001

    authors: Yao C,Luo J,Hsiao CH,Donelson JE,Wilson ME

    更新日期:2007-07-01 00:00:00

  • Giardia lamblia: stable expression of green fluorescent protein mediated by giardiavirus.

    abstract::Giardia lamblia, an early diverging eukaryote that infects several species including humans and a major agent of water-borne diarrhea throughout the world, can be infected with a double-stranded RNA virus, giardiavirus (GLV). A chimeric GLV cDNA and green fluorescent protein (GFP) according to the cis-acting signals o...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2004.12.006

    authors: Liu Q,Zhang X,Li J,Ying J,Chen L,Zhao Y,Wei F,Wu T

    更新日期:2005-03-01 00:00:00

  • Vaccination of sheep with purified cysteine proteinases of Fasciola hepatica decreases worm fecundity.

    abstract::There has been some evidence from several parasite systems that proteinases might have potential as protective antigens against parasitic infection. A cysteine proteinase complex identified in the regurgitant of adult F. hepatica was examined in this context. The thiol-cathepsin-related proteinases of M(r) 28,000 were...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1994.1014

    authors: Wijffels GL,Salvatore L,Dosen M,Waddington J,Wilson L,Thompson C,Campbell N,Sexton J,Wicker J,Bowen F

    更新日期:1994-03-01 00:00:00

  • Differentiation of Entamoeba histolytica: a possible role for enolase.

    abstract::The study of the encystation process of Entamoeba histolytica has been hampered by the lack of experimental means of inducing mature cysts in vitro. Previously we have found that cytoplasmic vesicles similar to the encystation vesicles of Entamoeba invadens are present in E. histolytica trophozoites only in amebas rec...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2011.05.001

    authors: Segovia-Gamboa NC,Talamás-Rohana P,Ángel-Martínez A,Cázares-Raga FE,González-Robles A,Hernández-Ramírez VI,Martínez-Palomo A,Chávez-Munguía B

    更新日期:2011-09-01 00:00:00

  • Giardia lamblia: interleukin 6 and tumor necrosis factor-alpha release from mast cells induced through an Ig-independent pathway.

    abstract::Giardia lamblia is a common cause of both acute and chronic diarrheal disease in humans worldwide. It has been shown that mast cells, IL-6 and TNF-alpha are substantially involved in the early control of G. lamblia infection in mice. However, no studies have yet been reported concerning the interaction between mast ce...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2010.06.013

    authors: Muñoz-Cruz S,Gómez-García A,Millán-Ibarra J,Giono-Cerezo S,Yépez-Mulia L

    更新日期:2010-11-01 00:00:00

  • Prevalence of Cryptosporidium species in recreational versus non-recreational water sources.

    abstract::Cryptosporidiosis, caused by the protozoan parasite Cryptosporidium, represents the major public health concern of water utilities in developed nations due to its small size, resistance to disinfection and ability to be shed in large numbers in faeces. In Australia, recreational access is not allowed on direct supply ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2012.04.015

    authors: Loganthan S,Yang R,Bath A,Gordon C,Ryan U

    更新日期:2012-08-01 00:00:00

  • Plasmodium falciparum and P. knowlesi: initial identification and characterization of malaria synthesized glycolipids.

    abstract::This is the first report establishing the existence of glycolipids synthesized by plasmodia, in particular Plasmodium falciparum. Trophozoites, schizonts, gametocytes, and gametes were metabolically labeled in vitro with [3H]glucosamine, [3H]galactose, [3H]glucose, [3H]mannose, [3H]fucose, [32P]inorganic phosphate, or...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(86)90016-0

    authors: Sherwood JA,Spitalnik SL,Aley SB,Quakyi IA,Howard RJ

    更新日期:1986-08-01 00:00:00

  • First report on isolation of Leishmania tropica from sandflies of a classical urban Cutaneous leishmaniasis focus in southern Iran.

    abstract::Shiraz district in south of Iran is a classical focus of Cutaneous leishmaniasis (CL) and previous research has consistently documented the etiologic agent to be Leishmania tropica and Leishmania major in urban and rural areas, respectively. However, none of the Phlebotomus sergenti, a known vector for L. tropica, of ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2010.05.020

    authors: Oshaghi MA,Rasolian M,Shirzadi MR,Mohtarami F,Doosti S

    更新日期:2010-12-01 00:00:00

  • Plasmodium falciparum: variations in the C-terminal cysteine-rich region of the merozoite surface protein-1 in field samples among Indian isolates.

    abstract::The cysteine-rich C-terminal region of the merozoite surface protein-1, MSP-119, of Plasmodium falciparum has been the most promising vaccine target antigen to date, based on protective immunization studies with recombinant proteins in mice and monkey models. To be further developed as a vaccine candidate, it is essen...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1999.4401

    authors: Lalitha PV,Malhotra P,Chattopadhyay R,Chauhan VS

    更新日期:1999-05-01 00:00:00

  • Susceptibility of clinical isolates of Leishmania aethiopica to miltefosine, paromomycin, amphotericin B and sodium stibogluconate using amastigote-macrophage in vitro model.

    abstract::Cutaneous Leishmaniasis (CL) caused by Leishmania aethiopica is a public health and social problem with a sequel of severe and mutilating skin lesions. It is manifested in three forms: localized CL (LCL), mucosal CL (MCL) and diffuse CL (DCL). Unresponsiveness to sodium stibogluconate (Sb(V)) is common in Ethiopian CL...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2013.01.022

    authors: Utaile M,Kassahun A,Abebe T,Hailu A

    更新日期:2013-05-01 00:00:00

  • Further characterization of refractoriness in Aedes aegypti (L.) to infection by Dirofilaria immitis (Leidy).

    abstract::Factors which control the expression of the refractory or susceptible condition to infection with Dirofilaria immitis in the mosquito. Aedes aegypti, were investigated using three protocols. (1) Microfilariae and prelarvae were injected into the hemocoel of susceptible A. aegypti. Some microfilariae and prelarvae deve...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(88)90057-4

    authors: Nayar JK,Knight JW,Bradley TJ

    更新日期:1988-06-01 00:00:00

  • Monocyte phagocytosis of malaria β-haematin in the presence of artemisinin, amodiaquine, chloroquine, doxycycline, primaquine, pyrimethamine and quinine.

    abstract::The intraerythrocytic malaria parasite digests haemoglobin to provide amino acids for metabolism and releases toxic haem that is sequestered into haemozoin, a non-toxic, insoluble, crystalline pigment. Following erythrocyte rupture, haemozoin is released into circulation and phagocytosed by monocytes. Phagocytosed hae...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2018.12.002

    authors: Cumming BM,Goldring JPD

    更新日期:2019-02-01 00:00:00

  • Effect of giardiasis combined with low-protein diet on intestinal absorption of glucose and electrolytes in gerbils.

    abstract::Studies have shown that symptomatic infection by Giardia lamblia causes acute or chronic diarrhea, dehydration, abdominal pain and malabsorption, leading to undernutrition and weight loss. The aim of the present study was to evaluate the effects of giardiasis and its combination with a low-protein diet on the intestin...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2012.04.016

    authors: Gomes MA,de Oliveira DR,de Freitas SE,de Pinho Viana M,Borges EL

    更新日期:2012-08-01 00:00:00

  • Trypanosoma evansi: a comparative study of four diagnostic techniques for trypanosomosis using rabbit as an experimental model.

    abstract::The goal of this study was to compare two parasitological diagnostic techniques, such as by Micro-Haematocrit Centrifugation Technique (MHCT) and Direct Microscopic Examination (DME) with a serological method (iELISA), and a molecular procedure PCR, in rabbits experimentally infected with Trypanosoma evansi, in order ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2011.02.010

    authors: Ramírez-Iglesias JR,Eleizalde MC,Gómez-Piñeres E,Mendoza M

    更新日期:2011-05-01 00:00:00

  • Schistosoma mansoni: surface membrane stability in vitro and in vivo.

    abstract::The human complement component C3b is known to bind in vitro to the surfaces of all developmental stages of schistosomes as a consequence of complement activation by the alternative pathway. C3b bound to Schistosoma mansoni parasites has now been used in combination with fluorescent labeled antibodies against C3b to l...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(86)90027-5

    authors: Ruppel A,McLaren DJ

    更新日期:1986-10-01 00:00:00

  • Toxoplasma gondii: purification of zoites from peritoneal exudates by eight methods.

    abstract::Many methods have been proposed for removing contaminating host cells from mouse peritoneal exudates infected with Toxoplasma gondii tachyzoites. Of these, eight established methods were compared. They were density gradients, sonication and trypsin digestion, differential centrifugation, haemolysin digestion, filtrati...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(84)90080-8

    authors: Dempster RP

    更新日期:1984-04-01 00:00:00

  • Plasmodium falciparum: induction of resistance to mefloquine in cloned strains by continuous drug exposure in vitro.

    abstract::A genetically homogeneous population of Plasmodium falciparum prepared by a single erythrocyte micromanipulation technique was used to produce lines of P. falciparum resistant to mefloquine hydrochloride in vitro. Parasites were maintained in a culture medium containing gradually increased concentrations of mefloquine...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(88)90082-3

    authors: Oduola AM,Milhous WK,Weatherly NF,Bowdre JH,Desjardins RE

    更新日期:1988-12-01 00:00:00

  • A new species and record of branchial parasitic isopods (Crustacea: Isopoda: Bopyridae: Pseudioninae) of porcellanid crabs from the Philippines.

    abstract::Branchial bopyrids infesting porcellanid crabs from the Philippines were investigated based on intertidal collections made in 1999-2000. Crabs of the genus Petrolisthes collected from sites in the northern Philippines were examined and two parasite species were found. One new pseudionine species found infesting Petrol...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2009.09.012

    authors: Williams JD,Madad AZ

    更新日期:2010-05-01 00:00:00

  • Behavioral and hormonal changes associated with the infective dose in experimental taeniasis in golden hamsters (Mesocricetus auratus).

    abstract:INTRODUCTION:It has been reported that behavioral changes relate to infection in different parasitoses. However, the relation between the extent of the behavioral changes and the magnitude of the infection has been scarcely studied. The aim of this study was to evaluate the relationship between different doses of infec...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2016.04.013

    authors: Domínguez-Roldan R,Hallal-Calleros C,Sciutto E,Hernández M,Aguirre-Flores V,García-Jiménez S,Báez-Saldaña A,Flores-Pérez FI

    更新日期:2016-07-01 00:00:00

  • Regulation of immunity in Trypanosoma cruzi infection.

    abstract::Immunity to T. cruzi is complex, involving among other components, antibody production, CD4+ helper cells, CD8+ T cells as both regulators and effectors of immunity, and possibly, double-negative T cells. In addition, several of these components have been implicated in pathogenesis in the chronic infection. Although t...

    journal_title:Experimental parasitology

    pub_type: 杂志文章,评审

    doi:10.1016/0014-4894(91)90013-m

    authors: Tarleton RL

    更新日期:1991-07-01 00:00:00