Evaluation of three enzyme immunoassays and a nucleic acid amplification test for the diagnosis of Clostridium difficile-associated diarrhea at a university hospital in Brazil

Silva, Rodrigo; Vilela, Eduardo García; Neves, Monique Silva; Lobato, Francisco Carlos Faria

doi:10.1590/0037-8682-0100-2014

Cited by 17 publications

(9 citation statements)

References 27 publications

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“…Seventy-six C. difficile isolates from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf isolated in between 2008 and 2015 were included ( Table I ). Human samples were collected from inpatients with suspicious CDI from the University Hospital of the Federal University of Minas Gerais (UFMG) ( Silva et al 2014c ). The samples from diarrhoeic dogs, foals, calves, and maned wolf ( Chrysocyon brachyurus ) were obtained directly from the rectum, at the Veterinary Hospital of UFMG at the time of the consultation, and were only collected from animals for which the main motivation for the consultation was the occurrence of diarrhoea ( Silva et al 2013a , b ).…”

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confidence: 99%

Clostridium difficile ribotypes in humans and animals in Brazil

Silva

Rupnik

Diniz

et al. 2015

Mem. Inst. Oswaldo Cruz

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Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies aboutC. difficile polymerase chain reaction (PCR) ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficileribotypes isolated from humans and animals in Brazil. Seventy-six C. difficile strains isolated from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf were distributed into 24 different PCR ribotypes. Among toxigenic strains, PCR ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4%) and eight (10.5%) samples, respectively. Fourteen different PCR ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.

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confidence: 99%

Clostridium difficile ribotypes in humans and animals in Brazil

Silva

Rupnik

Diniz

et al. 2015

Mem. Inst. Oswaldo Cruz

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“…This test provides a minimally instrumented, simple-to-interpret, fast diagnostic for BPV infection appropriate for a limited-resource setting [ 55 , 56 , 57 ]. The development of nucleic acid amplification tests (NAATs) similar to that used in the present study enhances both the sensitivity and specificity of diagnostics of various infectious agents compared to simple immunoassays [ 58 , 59 ]. In addition, the isothermal RPA method obviates the need for an expensive thermal cycler unit as needed for PCR.…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization and Developing a Point-of-Need Molecular Test for Diagnosis of Bovine Papillomavirus (BPV) Type 1 in Cattle from Egypt

El‐Tholoth

Mauk

Elnaker

et al. 2020

Animals

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Bovine papillomatosis is a viral disease of cattle causing cutaneous warts. A diagnosis of this viral infection is very mandatory for combating the resulting economic losses. Given the limited data available about bovine papillomavirus (BPV) in Egypt, the present study involved the molecular diagnosis of bovine papillomavirus type-1 (BPV-1), -2, -4, -5, and -10 in cattle presenting cutaneous warts on the head and neck from New Valley Province, Egypt. The phylogenetic analysis of the detected types of BPV was also performed, followed by developing a point-of-need molecular assay for the rapid identification of identified BPV types. In this regard, a total of 308 cattle from private farms in Egypt were clinically examined, of which 13 animals presented cutaneous warts due to suspected BPV infection. The symptomatic animals were treated surgically, and biopsies from skin lesions were collected for BPV-1, -2, -4, -5, and -10 molecular identification using polymerase chain reaction (PCR). The presence of BPV-1 DNA was confirmed in 11 collected samples (84.6%), while BPV-2, -4, -5, and -10 were not detected. Sequencing of the PCR products suggested the Egyptian virus is closely related to BPV found in India. An isothermal nucleic acid amplification test (NAAT) with labeled primers specific for the BPV-1 L1 gene sequence, and based on recombinase polymerase amplification (RPA), in combination with a lateral flow strip assay for the detection of RPA products, was developed and tested. The point-of-need molecular assay demonstrated a diagnostic utility comparable to PCR-based testing. Taken together, the present study provides interesting molecular data related to the occurrence of BPV-1 in Egypt and reveals the genetic relatedness of the Egyptian BPV-1 with BPV-1 found in buffalo in India. In addition, a simple, low-cost combined test was also validated for diagnosis of the infection. The present study suggests the necessity of future investigations about the circulating strains of the virus among the cattle in Egypt to assess their genetic relatedness and better understand the epidemiological pattern of the disease.

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“…In Brazil, EIA for detection of C. difficile toxins A and B in feces remains the most widely used technique for CDI diagnosis, because it is rapid, cheap, and easily performed. However, Silva et al have recently shown that all three commercial EIAs available in Brazil present low sensitivity, ranging from 61% to 68%, compared with the golden standard methods . Considering that roughly 75% of the health care‐associated diarrhea is related to causes other than CDI, the accurate diagnosis is of paramount importance …”

Section: Introductionmentioning

confidence: 99%