A multiplex PCR for unequivocal differentiation of all encapsulated and non-encapsulated genotypes of Trichinella

Zarlenga, Dante S.; Chute, M. B.; Martin, A; Kapel, Christian M. O.

doi:10.1016/s0020-7519(99)00107-1

Cited by 269 publications

(181 citation statements)

References 32 publications

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“…Trichinella larvae were identified at species level by multiplex polymerase chain reaction (multiplex PCR) according to Zarlenga et al (1999). Muscle larvae of T. spiralis (ISS003), T. nativa (ISS042), T. britovi (ISS002), and T. pseudospiralis (ISS013) reference strains were used as the controls.…”

Section: Trichinella Britovi Marten Badger Pcr Polandmentioning

confidence: 99%

Molecular identification of Trichinella britovi in martens (Martes martes) and badgers (Meles meles); new host records in Poland

Moskwa

Goździk

Bień

et al. 2012

Acta Parasitologica

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Trichinella larvae were detected in a marten (Martes martes) and a badger (Meles meles) in Poland. The animals were found dead following car accidents. All examined animals derived from the Mazurian Lake district, north-east Poland, near the village Kosewo Górne where Trichinella infection were earlier confirmed in wildlife; red foxes and wild boars. The muscle samples were examined by artificial pepsin-HCl digestion method. The parasites were identified as Trichinella britovi by multiplex polymerase chain reaction method. Larvae were found in two out of three martens and one out of seven examined badgers. This is the first report of the identification of Trichinella britovi larvae from martens and badgers in Poland.

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Section: Trichinella Britovi Marten Badger Pcr Polandmentioning

confidence: 99%

Molecular identification of Trichinella britovi in martens (Martes martes) and badgers (Meles meles); new host records in Poland

Moskwa

Goździk

Bień

et al. 2012

Acta Parasitologica

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“…With the same samples, the multiplex PCR produced bands for which both molecular weight and R: RAPD-PCR . M: Multiplex PCR (Zarlenga et al, 1999). *: Number of positive samples.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of two PCR-based techniques for molecular epidemiology in Finland, a high-endemic area with four sympatricTrichinellaspecies

Kapel¹,

Oivanen

Rosa

et al. 2001

Parasite

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Summary:Trichinella larvae collected from wildlife, domestic and synanthropic animals in Finland were identified to species by two molecular techniques: Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) and the recently described multiplex PCR. The RAPD-PCR was very sensitive to the sub-optimal preservation muscle larvae and resulting in weak and smeared bands on the gels for such material. However, the same samples yielded easily recognizable bands in the multiplex PCR; this latter technique is then recommended for epidemiological studies, especially when the preservation of the samples is sub-optimal. For larvae in good condition the unequivocal bands obtained by multiplex was the easiest identifiable. this is the first record of a mixed infection between T. spiralis and T. nativa in a naturally infected host. Raccoon dogs were the only host species from which all of the four Trichinella species were detected. Trichinella spiralis was found in both domestic animals and wildlife, but none of the sylvatic Trichinella species were detected in domestic pig. Because of the exceptionally high regional prevalence in wildlife (Oivanen & Oksanen, 1994;Oksanen et al, 1998) and recurrent infections in domestic swine (Oivanen & Oksanen, 1994), southern Finland presents an ideal opportunity for studies on the molecular epidemiology of Trichinella.The aims of the present study were to evaluate two techniques for molecular epidemiology in an area with sympatric species of Trichinella. It is the first time this multiplex PCR technique is used for an epidemiological study. MATERIALS AND METHODSM uscle samples were collected in Southern Finland (1993Finland ( -1997 from nine host species (bears, wolves, foxes, lynx, raccoon dogs, wild boars, domestic pigs, rats and cats). After initial examination, 87 muscle samples positive for Trichinella were stored at -20° C. However, all samples from rats were used for other purposes prior to the present study and were therefore thawed twice.Muscle larvae were released by HC1 pepsin digestion and recovered larvae were stored in 70 % ethyl-alcohol and stored at -20° C. Prior to molecular analysis, larvae were rehydrated in a series of alcohol and single larvae were finally placed in 5 µl sterile water.The condition of the larvae, was determined from the appearance of the larvae. Coiled and partly uncoiled larvae were described as in moderate condition and

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“…Ten pigs were experimentally infected with T. spiralis muscle larvae isolated by the digestion procedure from a naturally infected swine. T. spiralis larvae were identified at the species level by multiplex polymerase chain reaction (multiplex PCR) according to Zarlenga et al (1999). The counted T. spiralis ML exhibiting motility were suspended in 20% gelatin blocks.…”

Section: Hosts and Parasite Materialsmentioning

confidence: 99%

Detection of Experimental Swine Trichinellosis Using Commercial Elisa Test

Gondek¹,

Bień²,

Nowakowski³

2017

Polish Journal of Veterinary Sciences

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The aim of the study carried out on ten young (10-week old) pigs of the native Polish Large White breed experimentally infected with a low dose of 300 invasive muscle larvae (ML) of Trichinella spiralis was intravital detection of trichinellosis using the E-S ELISA test, determination of a variation level of IgG antibodies against excretory-secretory (E-S) antigens of T. spiralis muscle larvae and finally, describing the intensity of T. spiralis larvae infection in selected muscles. The pig sera were collected at 7 and 9 days prior to the experimental infection with T. spiralis and at 9, 14, 20, 23, 25, 27, 30, 33, 37, 41, 46 days post-infection (d.p.i.). The anti-T. spiralis IgG antibodies were detected by a commercial E-S ELISA test (PrioCHECK Trichinella Ab). Average intensity of the T. spiralis infection in the examined muscles of pigs ranged from 1.52 up to 43.09 larvae/g. The studies revealed that the E-S antigen in the ELISA test did not show cross-reaction with the sera of pigs infected with Oesophagostomum spp. The ELISA assay did not recognize trichinellosis in pigs until 27 days after the T. spiralis infection. The anti-T. spiralis IgG antibodies were first detected on day 30 post-infection. A statistically significant increase of IgG antibodies against T. spiralis ML E-S antigens was first observed between days 27-30 (p<0.01) post-infection, and a further significant rise in the antibody level occurred between days 27 and 33 (p<0.01); 30 and 33 (p<0.01); 33 and 37 (p<0.05) following infection.

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A multiplex PCR for unequivocal differentiation of all encapsulated and non-encapsulated genotypes of Trichinella

Cited by 269 publications

References 32 publications

Molecular identification of Trichinella britovi in martens (Martes martes) and badgers (Meles meles); new host records in Poland

Molecular identification of Trichinella britovi in martens (Martes martes) and badgers (Meles meles); new host records in Poland

Evaluation of two PCR-based techniques for molecular epidemiology in Finland, a high-endemic area with four sympatricTrichinellaspecies

Detection of Experimental Swine Trichinellosis Using Commercial Elisa Test

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