A novel one-step amplification refractory mutation system PCR (ARMS-PCR) for differentiation of canine parvovirus-2 variants

Dema, Anusha; Ganji, Vishweshwar Kumar; Yella, Narasimha Reddy; Putty, Kalyani

doi:10.1007/s11262-021-01861-w

Cited by 8 publications

(7 citation statements)

References 41 publications

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“…Typing of CAdV isolates as CAdV-1 or CAdV-2 was done based on differences in the melting temperature of the PCR product [ 32 ]. Typing for CPV-2/2a/2b/2c was done using a modification of a previously reported TARMS-PCR procedure [ 36 , 37 ].…”

Section: Methodsmentioning

confidence: 99%

A comprehensive molecular survey of viral pathogens associated with canine gastroenteritis

et al. 2023

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Viral pathogens are the primary cause of canine gastroenteritis. However, few structured comprehensive studies on the viral etiology of canine gastroenteritis have been conducted. In this study, 475 rectal swabs collected over three years (2018-2021) from clinical canine gastroenteritis cases were screened for the presence of six major enteric viruses – canine parvovirus 2 (CPV-2), canine distemper virus (CDV), canine adenovirus 2 (CAdV-2), canine coronavirus (CCoV), canine astrovirus (CaAstV), and canine rotavirus (CRV) – by real-time PCR. The most frequently detected virus was CPV-2, which was present in 64.8% of the samples (subtype 2a, 21.1%; 2b, 77.4%; 2c, 1.5%), followed by CDV (8%), CaAstV (7.2%), CCoV (5.9%), and CAdV-2 (4.6%). Two to four of these viruses in different combinations were found in 16.8% of the samples, and CRV was not detected. The complete genome sequences of Indian isolates of CDV, CCoV, and CaAstV were determined for the first time, and phylogenetic analysis was performed. This study highlights the need for routine prophylactic vaccination with the appropriate vaccines. Notably, 70.3% of animals vaccinated with DHPPiL were found to be positive for at least one virus. Hence, regular molecular analysis of the prevalent viruses is crucial for addressing vaccination failures. Supplementary Information The online version contains supplementary material available at 10.1007/s00705-022-05674-6.

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Section: Methodsmentioning

confidence: 99%

A comprehensive molecular survey of viral pathogens associated with canine gastroenteritis

et al. 2023

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Section: Nucleotide Sequencementioning

confidence: 98%

“…All the 166 samples were screened for ARMS -PCR to detect CPV-2 variants in cats using two pairs of primers (outer forward and reverse primers and inner forward and reverse primers) (Kalyani et al, 2021) (Table 1). The ARMS-PCR mix composition consist of (20ul) PCR master Mix (2x) 10 l, Forward primer-outer 0.6 ul, Reverse primer-outer 0.6 l, Forward primer-Inner 0.6 ul, Reverse primer-Inner 0.6 l , Template DNA 2 ul, Nuclease free water 5.6 l.…”

Section: Arms Pcrmentioning

confidence: 99%

“…Out of 166 faecal samples, 50 samples were shown positive in ARMS-PCR (Kalyani et al, 2021). ARMS-PCR was highly sensitive for detection and typing of canine parvovirus variant in cats (Kalyani et al, 2021). The outer primers targeting the conserved region of VP2 gene amplifies 631 bp in all antigenic variants of CPV-2 (Chander et al, 2016).…”

Section: Nucleotide Sequencementioning

confidence: 99%

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Molecular Characterization of Canine Parvo Virus Variant-2 in Cats in Chennai by Amplification-Refractory Mutation System Polymerase Chain Reaction

Harikrishnan¹,

Bharathi²,

Tirumurugaan³

et al. 2023

IJAR

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Background: Feline panleukopenia virus and canine parvovirus infections are highly contagious and serious enteric diseases of cats and dogs with high fatality rate. Canine parvo viral enteritis (CPV) is caused by CPV-2 antigenic variants (CPV-2a, 2b, 2c) is frequently reported in dogs worldwide leading to morbidity and mortality Infection in cats by canine parvo virus variants causes clinical signs similar to feline panleukopenia virus. CPV-2 variants have recently acquired the feline host rage, allowing it to infect both cats and dogs. Feline Panleukopenia virus is not the only parvovirus species affecting cats, in addition to Mink enteritis virus, the new variants of canine parvovirus, CPV-2a, 2b and 2c have also penetrated the feline host-range, and able to infect and replicate in cats, causing diseases indistinguishable from feline panleukopenia. The present study was taken to identify the canine parvovirus variants among domestic cats in Chennai and its role in transmission of CPV-2 between dogs through Amplification refractory mutation system. These findings suggest that species jump of CPV from dog to cats as well as CPV had presumably started a new process of readapting in feline hosts and confirmed the importance of viral host switching as a mechanism for the emergence of new viruses. Methods: In this study around 166 cat faecal swabs were collected during 2020-2022 in and around Chennai which were brought with the clinical signs of vomition and diarrhoea. Collected faecal samples were subjected to DNA extraction and Amplification Refractory Mutation System Polymerase Chain Reaction and 50 cat samples showed positivity towards canine parvovirus infection. Result: Of 166 faecal samples, 50 samples showed positive for the CPV-2a variant. By ARMS-PCR In this case, the 50 samples responding to outer forward and inner reverse primers and yielding a 492 bp amplicons in addition to the 631 bp common outer product as CPV-2a/2. This study suggested that CPV infection in cat showed that CPV has started a new process of readaptation in the feline host and confirming the importance of viral host switching mechanism as a mechanism for the emergence of new viruses. The introduction of CPV into the feline population raises concern about the efficacy of FPV-based vaccines in preventing CPV infection and point out the necessity for intensifying surveillance of parvovirus infection in cats.

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“…The antigenic typing of CAdV was done into CAdV-1 and CAdV-2 based on the difference in melting temperature (Balboni et al 2015). The antigenic typing for CPV-2/2a/2b/2c was done by a modi ed version of TARMS-PCR reported before (Dema et al 2021). Isolation Of Virus In Cell Lines DMEM containing 1% FBS was used for maintenance of Madin-Darby Canine Kidney (MDCK), A-72 (Canine broblast) or Epstein-Barr virus-transformed marmoset B lymphoblastoid (B95a) cell lines at 37 o C under 5% CO 2 .…”

Section: Screening Of Clinical Samples For Viral Etiology By Real Time Pcrmentioning

confidence: 99%

A Comprehensive Molecular Survey of Viral Pathogens Associated with Canine Gastroenteritis During 2018-21

Dema

Tallapally

Ganji

et al. 2022

Preprint

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Viral pathogens account for major aetiology of canine gastroenteritis in India. However, till date, there was no structured comprehensive study reported on the viral aetiology of canine gastroenteritis in India. To elucidate this, 475 rectal swabs collected over a period of three years (2018-2021) from clinical canine gastroenteritis cases were screened by real-time PCR for the presence of six majorly known enteric viruses (Canine parvovirus -2 (CPV-2), canine distemper virus (CDV), canine adenovirus-2 (CAdV-2), canine corona virus (CCoV), canine astrovirus (CaAstV), canine rotavirus (CRV)). 71.6% of the samples were found to harbour at least one of the five viruses tested (CPV-2/2a/2b/2c, CDV, CAdV-2, CCoV and CaAstV), whereas, there was no evidence of CRV. The overall incidence rate for each virus was found highest at 64.8% for CPV-2/2a/2b/2c (2a: 21.1%, 2b: 77.4%, 2c: 1.5%), followed by 8% for CDV, 7.2% for CaAstV, 5.9% for CCoV and 4.6% for CAdV-2 with 16.8% incidence for coinfections ranging from two to four viruses in different combinations. Further, for the first time we report the whole genome sequences of CDV, CCoV and CaAstV Indian isolates from canine gastroenteritis cases. Phylogenetic analysis of whole genome revealed that CDV clustered with Asia-1 lineage, CaAstV clustered with group III that consists of China and Brazil isolates, and CCoV clustered with group II that consists of China and Taipei isolates. This study adds to our knowledge on the prevalence of these pathogens in the Indian subcontinent and highlights the need for relevant and routine vaccine prophylactic measures. It is alarming to note that 70.3% of animals vaccinated with DHPPIL were found positive for at least one virus. Hence, regular molecular analysis of the prevalent viruses is crucial to address vaccination failures.

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A novel one-step amplification refractory mutation system PCR (ARMS-PCR) for differentiation of canine parvovirus-2 variants

Cited by 8 publications

References 41 publications

A comprehensive molecular survey of viral pathogens associated with canine gastroenteritis

A comprehensive molecular survey of viral pathogens associated with canine gastroenteritis

Molecular Characterization of Canine Parvo Virus Variant-2 in Cats in Chennai by Amplification-Refractory Mutation System Polymerase Chain Reaction

A Comprehensive Molecular Survey of Viral Pathogens Associated with Canine Gastroenteritis During 2018-21

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