Accuracy of the Kato-Katz method and formalin-ether concentration technique for the diagnosis of Clonorchis sinensis, and implication for assessing drug efficacy

Qian, Men-Bao; Yap, Peiling; Yang, Yue; Hai, Liang; Jiang, Zhihua; Li, Wei; Utzinger, Jürg; Zhou, Xiao‐Nong; Keiser, Jennifer

doi:10.1186/1756-3305-6-314

Cited by 36 publications

(26 citation statements)

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“…Recently we found that C. sinensis were able to mature and release eggs in BALB/c mice [ 9 ]. In this study, fecal examination was also performed by modified Kato-Katz egg counting technique [ 9 , 29 ] to further confirm the egg production capacities of the worms. However, no eggs were found in the faeces from both TLR2 wild-type and mutant mice after the infection.…”

Section: Discussionmentioning

confidence: 99%

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TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

et al. 2017

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BackgroundAlthough the responses of inducible nitric oxide synthase (iNOS) and associated cytokine after Clonorchis sinensis infection have been studied recently, their mechanisms remain incompletely understood. In this study, we investigated the effects of toll-like receptor 2 (TLR2) signals on iNOS/nitric oxide (NO) responses after C. sinensis infection. We also evaluated the correlations between iNOS responses and worm development, which are possibly regulated by TLR2 signal.MethodsTLR2 wild-type and mutant C57BL/6 J mice were infected with 60 C. sinensis metacercariae, and the samples were collected at 30, 60, 90 and 120 days post-infection (dpi). The total serum NO levels were detected using Griess reagent after nitrate was reduced to nitrite. Hepatic tissue samples from the infected mice were sliced and stained with hematoxylin and eosin (HE) to observe worm development in the intrahepatic bile ducts. The iNOS mRNA transcripts in the splenocytes were examined by real time reverse transcriptase polymerase chain reaction (qRT-PCR), and iNOS expression was detected by immunohistochemistry.ResultsDeveloping C. sinensis juvenile worms were more abundant in the intrahepatic bile ducts of TLR2 mutant mice than those of TLR2 wild-type mice. However, no eggs were found in the faeces of both mice samples. The serum levels of total NO significantly increased in TLR2 mutant mice infected with C. sinensis at 30 (t (5) = 2.595, P = 0.049), 60 (t (5) = 7.838, P = 0.001) and 90 dpi (t (5) = 3.032, P = 0.029). Meanwhile, no changes occurred in TLR2 wild-type mice compared with uninfected controls during the experiment. The iNOS expression in splenocytes showed unexpected higher background levels in TLR2 mutant mice than those in TLR2 wild-type mice. Furthermore, the iNOS mRNA transcripts in splenocytes were significantly increased in the TLR2 wild-type mice infected with C. sinensis at 30 (t (5) = 5.139, P = 0.004), 60 (t (5) = 6.138, P = 0.002) and 90 dpi (t (5) = 6.332, P = 0.001). However, the rising of iNOS transcripts dropped under the uninfected control level in the TLR2 mutant mice at 120 dpi (t (5) = -9.082, P < 0.0001). Both total NO and iNOS transcripts were significantly higher in the TLR2 mutant mice than those in the TLR2 wild-type mice at 30 (t (5) = 3.091/2.933, P = 0.027/0.033) and 60 dpi (t (5) = 2.667/6.331, P = 0.044/0.001), respectively. In addition, the remarkable increase of iNOS expressions was immunohistochemically detected in the splenic serial sections of TLR2 wild-type mice at 30 and 60 dpi. However, the expressions of iNOS were remarkably decreased in the splenocytes of both TLR2 wild-type and mutant mice at 120 dpi.ConclusionsThese results demonstrate that TLR2 signal plays an important role in the regulation of iNOS expression after C. sinensis infection. TLR2 signal is also beneficial to limiting worm growth and development and contributing to the susceptibility to C. sinensis in which the iNOS/NO reactions possibly participate.Electronic supplementary materialThe online version of this articl...

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Section: Discussionmentioning

confidence: 99%

“…Stool samples were collected from each infection group every other day and were mixed together every 20 days. Eggs were counted by the Kato-Katz egg counting technique [ 29 ]. For perfect blending, 1 ml of pure water was added into 1 g of stool of each sample.…”

Section: Methodsmentioning

confidence: 99%

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

et al. 2017

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“…The stool samples were thoroughly mixed after complete humidification. Triplicate Kato-Katz thick smears using standard 41.7 mg templates [ 24 ] were prepared from each stool sample. The number of C. sinensis eggs was counted and presented as the geometric mean of eggs per 1 g of stool (GM EPG).…”

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Pathological Lesions and Inducible Nitric Oxide Synthase Expressions in the Liver of Mice Experimentally Infected with <i>Clonorchis sinensis</i>

et al. 2015

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The nitric oxide (NO) formation and intrinsic nitrosation may be involved in the possible mechanisms of liver fluke-associated carcinogenesis. We still do not know much about the responses of inducible NO synthase (iNOS) induced by Clonorchis sinensis infection. This study was conducted to explore the pathological lesions and iNOS expressions in the liver of mice with different infection intensity levels of C. sinensis. Extensive periductal inflammatory cell infiltration, bile duct hyperplasia, and fibrosis were commonly observed during the infection. The different pathological responses in liver tissues strongly correlated with the infection intensity of C. sinensis. Massive acute spotty necrosis occurred in the liver parenchyma after a severe infection. The iNOS activity in liver tissues increased, and iNOS-expressing cells with morphological differences were observed after a moderate or severe infection. The iNOS-expressing cells in liver tissues had multiple origins.

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“…Contradictory results have been reported in the literature on the effectiveness of Kato-Katz and Formolether techniques in the detection of gastrointestinal parasites in faeces. Some authors have noted that the Kato-Katz technique was better and significantly more sensitive than the other method (18)(19) while others have reported that the Formol-ether technique had a better sensitivity than the Kato-Katz method (20)(21). According to Knight et al, (22), the concentration of eggs in the faeces enabled the detection of more patients infected with S. mansoni when the Formol-ether technique was used.…”

Section: Discussionmentioning

confidence: 99%

Comparison of two diagnostic techniques to determine the prevalence of <i>Schistosoma mansoni</i> infections in Cameroonian school children

Soppa¹,

Nkengni²,

Nguepnang³

et al. 2019

Af J Clin Exp Micro

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Background: The Kato-Katz technique is recommended for diagnosis of Schistosoma mansoni infection by the World Health Organization. However, egg counts are subject to variability. The aim of this study is to determine the prevalence of S. mansoni infection in school children using two different techniques and to recommend the technique that should be routinely used in the diagnosis of this infection. Methodology: Field investigations on faecal samples from 299 Cameroonian school children were carried out in 2016 to compare the effectiveness of the Kato-Katz and Formalin-ether techniques in diagnosis of S. mansoni infections. Results: Schistosome eggs were detected in 37 (12.3%) samples with the Kato-Katz technique and 61 (20.4%) samples with the Formalin-ether technique. The difference between the prevalence observed for the two techniques was significant in males and age group 10-12 years (p < 0.5). Conclusion: The Formalin-ether technique was more sensitive than the Kato-Katz method for detecting S. mansoni eggs in faecal matter. Despite its cost, the Formalin-ether technique can be routinely used in the laboratory for epidemiological studies of intestinal schistosomiasis

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Accuracy of the Kato-Katz method and formalin-ether concentration technique for the diagnosis of Clonorchis sinensis, and implication for assessing drug efficacy

Cited by 36 publications

References 35 publications

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

Pathological Lesions and Inducible Nitric Oxide Synthase Expressions in the Liver of Mice Experimentally Infected with <i>Clonorchis sinensis</i>

Comparison of two diagnostic techniques to determine the prevalence of <i>Schistosoma mansoni</i> infections in Cameroonian school children

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Accuracy of the Kato-Katz method and formalin-ether concentration technique for the diagnosis of Clonorchis sinensis, and implication for assessing drug efficacy

Cited by 36 publications

References 35 publications

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

Pathological Lesions and Inducible Nitric Oxide Synthase Expressions in the Liver of Mice Experimentally Infected with &lt;i&gt;Clonorchis sinensis&lt;/i&gt;

Comparison of two diagnostic techniques to determine the prevalence of <i>Schistosoma mansoni</i> infections in Cameroonian school children

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Pathological Lesions and Inducible Nitric Oxide Synthase Expressions in the Liver of Mice Experimentally Infected with <i>Clonorchis sinensis</i>