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Detection of Fasciola Hepatica in Lori Sheep Using Polymerase Chain Reaction and Conventional Diagnostic Methods in Western Iran.

ABSTRACT: Fascioliasis is an emerging and important food and water-borne disease in human communities which has become one of the most important health challenges in countries, like Iran. It causes weight loss, a decrease in feed conversion ratio as well as milk and meat production, and also reduces fertility in animals the prevalence of fasciolosis is increasing in some regions of the world due to various factors. Different methods have been used for the detection of Fasciola hepatica in animals. This study is the first to detect F. hepatica in Lori sheep using polymerase chain reaction (PCR) and conventional diagnostic methods in Western Iran. During three months, 195 fecal samples were collected from sheep in Lorestan province, Iran, using the stratified random sampling method. The conventional diagnostic methods, including wet mount microscopic examination and concentration assays, as well as the PCR technique targeting the intergenic spacer gene of F. hepatica, were used for the detection of the parasite in sheep. In total, 4 (2.1%) out of 195 examined stool samples were positive for F. hepatica based on the conventional assays. The PCR test was positive for F. hepatica in7 (3.6%) samples of 195 studied specimens. Statistical analyses of the data revealed that there is a significant difference between the results of diagnostic methods for F. hepatica detection (P=0.0421). Finally, the results showed that PCR has more diagnostic sensitivity, compared to conventional diagnostic methods, including the concentration techniques and microscopic examination. Hence, it can be advised to use PCR for the detection of F. hepatica in sheep.

SUBMITTER: Amiri S

PROVIDER: S-EPMC8410177 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Background and aimDifferent polymerase chain reaction (PCR) techniques have and are still being used for the direct detection of Brucella DNA in serum samples of different animal species and humans without being validated or properly validated, resulting in discrepancies. Thus, this study aimed to evaluate the diagnostic performance of the TaqMan Real-Time-PCR (RT-PCR) targeting the bcsp31 gene versus conventional PCR for the accurate diagnosis of brucellosis at the genus level in cattle sera.Materials and methodsOne hundred and eighty-four serum samples were collected from bacteriologically positive and negative cows with ages ranging from 1 to 5 years old at some infected private farms in the Nile Delta under quarantine measures as well as brucellosis free farms. These samples were classified into four groups after serological diagnosis and investigated by TaqMan RT-PCR and conventional PCR targeting the IS711 gene for Brucella DNA detection. The diagnostic performance characteristics of both PCR techniques were estimated considering the bacteriological results as a gold standard.ResultsTaqMan RT-PCR revealed superiority over conventional PCR; it was able to detect Brucella DNA in 95% (67/70) and 89% (25/28) of the cattle sera samples belonging to Group 1 (serologically and bacteriologically positive) and Group 2 (serologically negative but bacteriologically positive), respectively. On evaluating the diagnostic performance, TaqMan RT-PCR showed superior diagnostic sensitivity (93.9%), diagnostic specificity (88.4%), performance index (182.3), almost perfect kappa agreement (0.825±0.042), strong positive correlation (r=0.826), high accuracy based on the receiver operating characteristic (ROC) curve, and area under the ROC curve (0.911) at p<0.05 and CI of 95%.ConclusionA cattle serum sample is not the metric of choice for targeting Brucella genomic DNA by conventional PCR. The time-saving and rapid TaqMan RT-PCR method revealed a better diagnostic performance in the detection of Brucella DNA in cattle sera. Such performance offered by TaqMan RT-PCR may be considered a step toward the possibility of using such technology in the direct differentiation between Brucella-infected and -vaccinated cattle immunized by smooth vaccines from cattle sera using primers specific for such vaccines.

Dataset Information

Detection of Fasciola Hepatica in Lori Sheep Using Polymerase Chain Reaction and Conventional Diagnostic Methods in Western Iran.

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