Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia

Tan, Liping; Yu, Xianjun; Abdullahi, A. Y.; Wu, Shengde; Gong, Zheng; Hu, Wei; Song, Meiran; Wang, Zhen; Jiang, Biao; Li, Guoqing

doi:10.1007/s00436-015-4636-3

Cited by 13 publications

(6 citation statements)

References 24 publications

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“…Assemblages A and B have been reported in many mammals, including humans [ 12 – 15 ] and dogs [ 16 , 17 ] and have a zoonotic potential which renders them of great public health concern [ 9 , 18 – 20 ]. Currently, there is cumulative evidence that dogs act as a source of contamination for humans and pose a risk especially for pet owners and shelter staff [ 21 , 22 ]. Also, giardiosis outbreaks due to contaminated drinking water and food have been reported [ 23 – 26 ].…”

Section: Introductionmentioning

confidence: 99%

Rapid on-site diagnosis of canine giardiosis: time versus performance

et al. 2020

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Background Infections by protozoans of the genus Giardia are a common cause of diarrhea in dogs. Canine giardiosis constitutes a disease with a zoonotic potential; however, it is often underestimated due to its challenging diagnosis. The objective of the study was to assess the diagnostic performance of an immunochromatographic strip test (SpeedTMGiardia, Virbac, France) comparing it with microscopy (zinc sulfate flotation) by utilizing the combination of an enzyme immunoassay (ProSpecTTMGiardia EZ Microplate Assay, Oxoid Ltd., UK) and the PCR as the gold standard. A positive result in both ELISA and PCR was set as the gold standard. Methods Initially, fecal samples from dogs with clinical signs compatible with giardiosis were tested with the SpeedTMGiardia test and separated into two groups of 50 samples each: group A (positive) and group B (negative). Thereafter, all samples were examined by zinc sulfate centrifugal flotation technique and assayed by the ProSpecTTMGiardia Microplate Assay and PCR. The performance of the SpeedTMGiardia and zinc sulfate centrifugal flotation tests were calculated estimating sensitivity, specificity, and positive and negative likelihood ratio; the chi-square and McNemar tests were used for the comparison of the two methods. Results Giardia cysts were not detected by microscopy in 16 out of the 50 samples (32%) of group A and in none of group B samples. Eight out of 50 samples in group B (16%) were tested positive both with the ProSpecTTMGiardia Microplate Assay and PCR. Fecal examination with the SpeedTMGiardia test was more sensitive (86.2%) than the parasitological method (58.6%, P < 0.001) while the specificity of both methods was 100%. Conclusions The SpeedTMGiardia test is an easy-to-perform diagnostic method for the detection of Giardia spp., which can increase laboratory efficiency by reducing time and cost and decrease underdiagnosis of Giardia spp. infections. This immunochromatographic strip test may be routinely exploited when a rapid and reliable diagnosis is required, other diagnostic techniques are unavailable and microscopy expertise is inefficient. In negative dogs with compatible clinical signs of giardiosis, it is recommended either to repeat the exam or proceed with further ELISA and PCR testing.

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Section: Introductionmentioning

confidence: 99%

Rapid on-site diagnosis of canine giardiosis: time versus performance

et al. 2020

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“…To solve this problem, a variety of molecular methods, including nested PCR, PCR-SSCP, PCR-RFLP and multiplex PCR (Gasser et al ., 1993, 1998; Palmer et al ., 2007; Hu et al ., 2015), have been developed in recent years. However, they still have some disadvantages, such as being laborious; having poor specificity, with false positives occurring frequently in PCR-SSCP (Sheffield et al ., 1993); and having low sensitivity, with many false negatives in PCR-RFLP (Tan et al ., 2015; George et al ., 2016). Besides, the risk of contamination is increased in these methods because of complex operation steps.…”

Section: Discussionmentioning

confidence: 99%

“…To evaluate the established HRM method, a series of experiments on its stability, sensitivity and accuracy were performed using the above-mentioned quantitative PCR (qPCR)-HRM reaction system and the conditions, with few modifications, reported in our previous work (Tan et al ., 2015). Intra-assays were tested seven times, while inter-assays were tested three times for each plasmid.…”

Section: Methodsmentioning

confidence: 99%

“…No prior sequence information is needed to enable the detection of previously unknown, and even complex, sequence variations in a simple and straightforward way (Erali et al ., 2008). HRM has been widely used in the rapid identification and differentiation of many parasitic protozoans (Tan et al ., 2015) because of its high degree of automation, high throughput, closed-tube operation and low cost (Reed & Wittwer, 2004; Wilson et al ., 2005). In helminths, Areekit et al .…”

Section: Introductionmentioning

confidence: 99%

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Molecular differentiation of three canine and feline hookworms in South China through HRM analysis

et al. 2018

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To investigate the prevalence of canine and feline hookworms in South China, and to assess the risk of zoonotic hookworms to humans, one pair of primers (HRM-F/HRM-R) was designed to establish a high-resolution melting (HRM) method based on internal transcribed spacer 1 (ITS-1) rDNA for the detection of Ancylostoma ceylanicum, A. caninum and A. tubaeforme infection. The results showed that the HRM for the three hookworms produced different melting-curve profiles, where melting temperature (T m) values were 84.50°C for A. ceylanicum, 82.25°C for A. caninum and 81.73°C for A. tubaeforme, respectively. The reproducibility of intra- and inter-assay melting curves was almost perfect. The lowest concentration detected was about 5.69 ×10-4 g/μl. The HRM detection results from 18 canine and feline hookworm samples were in complete accordance with their sequencing results. The HRM method was more sensitive than the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in the detection of 98 clinical samples. It is concluded that the HRM method can differentiate between A. ceylanicum, A. caninum, A. tubaeforme and their mixed infections, which may provide important technical support for the zoonotic risk assessment and molecular epidemiological survey of canine and feline hookworms.

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“…On the other hand, China has new trends in the prevalence of parasitic diseases, including the increase of foodborne parasitic diseases (clonorchiasis, angiostrongyliasis, etc) and imported parasitic diseases (imported malaria, imported schistosomiasis, etc), as well as zoonotic diseases caused by pets (toxoplasmosis, cryptosporidiosis, etc). 7 Therefore, the teaching contents and methods of human parasitology have to be adjusted according to the epidemic characteristics of parasitic diseases. 7 , 8 In addition, during the traditional teaching process, teachers and students lack appropriate online platforms and tools for immediate interaction and learning feedback outside the classroom.…”

Section: Introductionmentioning

confidence: 99%

Development of the WeChat Public Account I Love Parasitology and its Preliminary Application in the Teaching of Human Parasitology

Zhu,

Deng,

Guan

et al. 2024

Journal of Medical Education and Curricular Development

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OBJECTIVE To better construct teaching resources, enhance real-time interaction and feedback between teachers and students in and out of class, and improve the teaching quality of parasitology, our team set up a WeChat public account I love Parasitology. METHODS The data sources were mainly from original pictures and multimedia materials of different parasites collected and produced by our team, as well as related materials collected from traditional publications and digital media. With the instant interactive platform, course schedules and corresponding teaching contents were sent by push notifications, case-based learning was carried out, and 2-way communication between students and teachers was achieved. Teaching effectiveness was assessed using a self-evaluation questionnaire. RESULTS A WeChat public account suitable for our daily teaching of parasitology was established. The second recursion and implementation of the learning resources allowed students to conduct in-depth reading and get unrestricted access to high-quality resources through the public account. In addition, all contents were in digital forms and made the original resources reborn, which would make up for our current and future shortage of physical teaching specimens. Moreover, the results from the questionnaire indicated that all these actions encouraged students to master theoretical knowledge, improved their abilities of case analysis and communication, and increased their knowledge of academic progress. CONCLUSION Our WeChat public account can provide excellent learning materials for students and is a good supplement to the routine education of human parasitology.

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Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia

Cited by 13 publications

References 24 publications

Rapid on-site diagnosis of canine giardiosis: time versus performance

Rapid on-site diagnosis of canine giardiosis: time versus performance

Molecular differentiation of three canine and feline hookworms in South China through HRM analysis

Development of the WeChat Public Account I Love Parasitology and its Preliminary Application in the Teaching of Human Parasitology

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