Highly sensitive multiplex PCR for simultaneous detection and discrimination of Dirofilaria immitis and Dirofilaria repens in canine peripheral blood

Gioia, G.; Lecová, Lenka; Genchi, Marco; Ferri, Emanuele; Genchi, C.; Mortarino, Michele

doi:10.1016/j.vetpar.2010.04.027

Cited by 70 publications

(49 citation statements)

References 13 publications

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“…genomic DNA was extracted from 100 µL of each positive sample using the DNeasy blood and tissue kit (Qiagen, Leipzig, Germany) following the manufacturer's instructions. An equimolar combination of general and specific primers (12SF/12SRdeg/12SF2B/12SR2) to amplify a portion of the small subunit ribosomal RNA gene of the mitochondrion (12S rDNA) were used to perform a multiplex polymerase chain reaction (PCR) according to Gioia et al (2010). The PCR products were detected on a 2.5% ethidium bromide-stained low-melting agarose gel (BIO-RAD, Madrid, Spain).…”

Section: Laboratory Techniquesmentioning

confidence: 99%

Geographic distribution of canine heartworm (Dirofilaria immitis) infection in stray dogs of eastern Romania

et al. 2016

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A survey was conducted in the eastern part of Romania to assess the prevalence and geographical distribution of Dirofilaria immitis in dogs. Plasma samples were collected from 458 stray dogs hosted in shelters in 8 counties and tested serologically for the presence of heartworm. In addition, 45 blood samples from dogs of a shelter in Galati city were examined by the modified Knott and multiplex polymerase chain reaction (PCR) techniques. The immmunological assay showed a heartworm infection prevalence of 8.9% in the dogs. Optical density results for positive samples ranged between 0.217 and 2.683. Geographical information systems (GIS) were used to produce overlays of distribution maps of D. immitis prevalence and predictive maps based on temperature suitability. High prevalence of D. immitis was found in the central East up to the northern border of the country, i.e.Galati county (60%), followed by the counties of Vaslui (12.0%) and Iasi (7.7%). Out of 45 samples examined using the Knott test, 23 were positive for circulating microfilariae (51.1%), while 19 dogs were positive for D. immitis and 4 for both D. immitis and D. repens with the multiplex PCR test. The high prevalence for D. immitis shown in dogs in the Southeast (Galati, 42.2%) also by multiplex PCR gave strong support to the results achieved by the serological tests. The present study confirms the ability of GIS to predict the distribution and epidemiology of dirofilariosis in different geographical territories as has been already demonstrated by the empirical epidemiological data obtained at the continental, national and intraregional levels.

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Section: Laboratory Techniquesmentioning

confidence: 99%

Geographic distribution of canine heartworm (Dirofilaria immitis) infection in stray dogs of eastern Romania

et al. 2016

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“…After macroscopic identification, both worms were stained with Horen's trichrome stain, and paraffin-embedded transverse sections were stained with hematoxylin and eosin for microscopic analysis. Finally, a multiplex polymerase chain reaction (PCR) assay and sequencing were performed for simultaneous detection and discrimination of D. immitis and D. repens using the protocol described in Gioia et al (2010).…”

Section: Human and Canine Case Reports: Morphological And Molecular Dmentioning

confidence: 99%

“…Based on the size, morphological features (Furtado et al 2010) and molecular analysis (Gioia et al 2010), the worms were identified as a female (canine case) and a male (human case) of D. immitis.…”

Section: Human and Canine Case Reports: Morphological And Molecular Dmentioning

confidence: 99%

Detection of Dirofilaria immitis in mid-western arid Argentina

Cuervo

Sierra

Waisman

et al. 2013

Acta Parasitologica

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Dirofilariosis, caused by Dirofilaria immitis and D. repens, is (re-) emerging worldwide. Dogs are the main reservoirs, while human infection has recently become an important focus of interest and attention. In Argentina, canine D. immitis infection has been described in eastern and northern subtropical and temperate humid regions, but never reported in mid-western arid regions so far. In this research note we report for the first time the occurrence of autochthonous human and canine D. immitis infection in the region.

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“…Death is mainly due to cachexia or respiratory distress after thromboembolism (10). Laboratory diagnosis of dirofilariasis involves the detection and identification of microfilaria in the peripheral blood (8,27), performance of immunological tests (13,20,23), and molecular diagnostic methods (5,9,26). If dogs manifest symptoms and the results of blood tests are negative, then radiography of the thorax should be performed (15).…”

Section: Introductionmentioning

confidence: 99%

Efficiency of modified therapeutic protocol in the treatment of some varieties of canine cardiovascular dirofilariasis

Stepanovic

Ilić

Krstić

et al. 2015

Bulletin of the Veterinary Institute in Pulawy

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The paper presents clinical diagnostic approaches and therapeutic effects of a specific protocol for the treatment of dogs with cardiovascular dirofilariasis in the Belgrade City (Serbia) territory. The study involved 50 privately owned dogs of different breeds, gender, and age, all showing signs of cardio -respiratory disorders. In addition to a general physical examination, blood tests were done to detect microfilaria and adult forms, and X-ray, ECG, and echocardiography were performed as well. At the first examination, 34 out of 50 examined dogs were positive for microfilaria and adult forms. Because of a lack of drug used as "the golden standard" in dirofilariasis treatment, it involved a combination of doxycycline (10 mg/kg) and ivermectin (6 µg/kg) supported with Advocate -Bayer spot-on. After six months, the first control was performed while continuing treatment with the aforesaid protocol, and the second control was performed after 12 months. Of the 34 treated dogs, all were negative for microfilaria, as early as after the first six months of the treatment (100%). One dog was positive for adult forms of the parasite after six and 12 months. In echocardiography and X-ray examination after 12 months, six dogs showed evident chronic changes. At controls conducted at sixth month and at one year, the implemented therapy was successful in 97.05% (33/34) of primarily infected dogs.

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Highly sensitive multiplex PCR for simultaneous detection and discrimination of Dirofilaria immitis and Dirofilaria repens in canine peripheral blood

Cited by 70 publications

References 13 publications

Geographic distribution of canine heartworm (Dirofilaria immitis) infection in stray dogs of eastern Romania

Geographic distribution of canine heartworm (Dirofilaria immitis) infection in stray dogs of eastern Romania

Detection of Dirofilaria immitis in mid-western arid Argentina

Efficiency of modified therapeutic protocol in the treatment of some varieties of canine cardiovascular dirofilariasis

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