• Parasites, Hosts and Diseases
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• 제51권1호
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• pp.93-98
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• 2013

A field applicable diagnostic technique, the dipstick assay, was evaluated for its sensitivity and specificity in diagnosing human Schistosoma mansoni infection. A monoclonal antibody (mAb) against S. mansoni adult worm tegumental antigen (AWTA) was employed in dipstick and sandwich ELISA for detection of circulating schistosome antigen (CSA) in both serum and urine samples. Based on clinical and parasitological examinations, 60 S. mansoni-infected patients, 30 patients infected with parasites other than schistosomiasis, and 30 uninfected healthy individuals were selected. The sensitivity and specificity of dipstick assay in urine samples were 86.7% and 90.0%, respectively, compared to 90.0% sensitivity and 91.7% specificity of sandwich ELISA. In serum samples, the sensitivity and specificity were 88.3% and 91.7% for dipstick assay vs. 91.7% and 95.0% for sandwich ELISA, respectively. The diagnostic efficacy of dipstick assay in urine and serum samples was 88.3% and 90.0%, while it was 90.8% and 93.3% for sandwich ELISA, respectively. The diagnostic indices of dipstick assay and ELISA either in serum or in urine were statistically comparable (P>0.05). In conclusion, the dipstick assay offers an alternative simple, rapid, non-invasive technique in detecting CSA or complement to stool examinations especially in field studies.

• Food Science and Biotechnology
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• 제15권6호
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• pp.990-992
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• 2006

In a previous study, we obtained polyclonal antibodies against the organophosphorus insecticide fenitrothion and developed an enzyme-linked immunosorbent assay (ELISA) for this pesticide. Using these antibodies and an enzyme tracer, a direct competitive ELISA method specific for fenitrothion using a dipstick format was developed. Dipstick ELISA using antibodies to fenitrothion immobilized on an Immunodyne membrane allowed the quick visual detection of fenitrothion at concentrations above $10\;{\mu}g/L$. The $IC_{50}$ value of dipstick ELISA using reflectance detection was $27\;{\mu}g/L$ with a detection limit of $2\;{\mu}g/L$. The recovery of fenitrothion from spiked lettuce and rice samples using the dipstick ELISA ranged from 87-107%.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1316-1319
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• 2006

An antibody displayed phage collection (SBAE-2R), screened from a human synthetic phage antibody library (Fab 21ox), was used for the determination of dextran. The dextran-binding affinity was determined by serologically specific transmission electron microscopy (TEM) and a paper dipstick assay. The phage collection was distributed over the dextrancoated grids with 39$\pm$25 phages/$\mu$m$^2$ on the grids. Phages were not seen on dextran-coated grids exposed to the Fab 2lox phage library. The phage collection (SBAE-2R) produced 54$\pm$3 color normalized intensity (N.I.) from 125 ppm to 1,000 ppm of dextran and 5$\pm$1 (N.I.) for 63 ppm of dextran in a paper dipstick assay. This research extends the analytical options for dextran analysis by antibody displayed phage with a minimum of equipment usage.

• Food Science and Biotechnology
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• 제14권6호
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• pp.743-746
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• 2005

A dipstick-type immunochemical biosensor for the detection of the organophosphorus insecticide fenthion was developed using a screen-printed electrode system as an amperometric transducer with polyclonal antibodies against fenthion as a bioreceptor. The assay of the biosensor involved competition between the pesticide in the sample and pesticide-glucose oxidase conjugate for binding to the antibody immobilized on the membrane. This was followed by measurement of the activity of the bound enzyme by the supply of the enzyme substrate (glucose) and amperometric determination of the enzyme reaction product ($H_2O_2$). The activity of the bound enzyme was inversely proportional to the concentration of pesticide. The optimized sensor system showed a linear response against the logarithm of the pesticide concentration ranging from $10^{-2}$ to $10^3\;{\mu}g/L$.

• Journal of Microbiology and Biotechnology
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• 제31권12호
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• pp.1672-1683
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• 2021

Vibrio parahaemolyticus is recognized as one of the most important foodborne pathogens responsible for gastroenteritis in humans. The bla_CARB-17 gene is an intrinsic β-lactamase gene and a novel species-specific genetic marker of V. parahaemolyticus. In this study, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) was developed targeting this bla_CARB-17 gene. The specificity of LAMP-LFD was ascertained by detecting V. parahaemolyticus ATCC 17802 and seven other non-V. parahaemolyticus strains. Finally, the practicability of LAMP-LFD was confirmed by detection with V. parahaemolyticus-contaminated samples and natural food samples. The results showed that the optimized reaction parameters of LAMP are as follows: 2.4 mmol/l Mg²⁺, 0.96 mmol/l dNTPs, 4.8 U Bst DNA polymerase, and an 8:1 ratio of inner primer to outer primer, at 63℃ for 40 min. The optimized reaction time of the LFD assay is 60 min. Cross-reactivity analysis with the seven non-V. parahaemolyticus strains showed that LAMP-LFD was exclusively specific for V. parahaemolyticus. The detection limit of LAMP-LFD for V. parahaemolyticus genomic DNA was 2.1 × 10^-4 ng/μl, corresponding to 630 fg/reaction and displaying a sensitivity that is 100-fold higher than that of conventional PCR. LAMP-LFD in a spiking study revealed a detection limit of approximately 6 CFU/ml, which was similar with conventional PCR. The developed LAMP-LFD specifically identified the 10 V. parahaemolyticus isolates from 30 seafood samples, suggesting that this LAMP-LFD may be a suitable diagnostic method for detecting V. parahaemolyticus in aquatic foods.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.454-464
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• 2019

Salmonellosis is a highly contagious bacterial disease that threatens both human and poultry health. Tests that can detect Salmonella in the field are urgently required to facilitate disease control and for epidemiological investigations. Here, we combined loop-mediated isothermal amplification (LAMP) with a chromatographic lateral flow dipstick (LFD) to rapidly and accurately detect Salmonella. LAMP primers were designed to target the Salmonella invA gene. LAMP conditions were optimized by adjusting the ratio of inner to outer primers, $MgSO_4$ concentration, dNTP mix concentration, amplification temperature, and amplification time. We evaluated the specificity of our novel LAMP-LFD method using six Salmonella species and six related non-Salmonella strains. All six of the Salmonella strains, but none of the non-Salmonella strains, were amplified. LAMP-LFD was sensitive enough to detect concentrations of Salmonella enterica subsp. enterica serovar Pullorum genomic DNA as low as $89fg/{\mu}l$, which is 1,000 times more sensitive than conventional PCR. When artificially contaminated feed samples were analyzed, LAMP-LFD was also more sensitive than PCR. Finally, LAMP-LFD gave no false positives across 350 chicken anal swabs. Therefore, our novel LAMP-LFD assay was highly sensitive, specific, convenient, and fast, making it a valuable tool for the early diagnosis and monitoring of Salmonella infection in chickens.

• Journal of Yeungnam Medical Science
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• 제8권1호
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• pp.42-52
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• 1991

최근 널리 사용되고 있는 제조회사가 다른 두가지 요시험지봉(Combur-9-test$^{(R)}$ RL & Multistix$^{(R)}$-SG)으로 요검사를 실시하면, 그 결과치의 표현에 있어 차이가 있으므로, 결과치 판정시 질병의 진단이나 치료의 효과 판정 및 경과관찰에 있어서 발생할 수 있는 혼란을 줄이고자, 환자검체와 표준검체에서 실시한 요시험지봉 검사결과를 비교 검토하여 아래와 같은 결론을 얻었다. 또한 두 시험지봉검사에 결과를 교환함에 있어 상응하는 결과치도 조사하였다. Combur-9-test RL 및 Multistix-SG를 이용하여, 1990년 12월 16일 부터 15일간 영남대학교 의과대학 부속병원 임상병리과에 요검사가 의뢰된 501명의 환자검체를 대상으로 하여 요시험지봉으로 화학적검사를 반정량적으로 시행한 결과 대체로 ${\pm}1$등급 범위내에서 일치하였다. Combur-9-test RL을 이용한 leukocyte esterase 검사와 요침사검사간의 비교에서 현미경적 농뇨와의 일치율은 83.7%, 민감도 48.1%, 특이도 90.3%, 양성예측율 47.4% 및 음성예측율 90.5%로 나타났다. Combur-9-test RL을 사용한 nitrite검사와 요배양검사간의 비교에서 요배양 양성과의 일치율은 93.0%, 민감도 19.4%, 특이도 84.7%, 양성예측율 53.8% 및 음성예측율 94.1%로 나타났다. 단백의 주 성분이 알부민 및 글로부린인 검체에서 실시한 요단백검사에서는 양자 모두에서 sulfosalicylic acid 법이 가장 민감하였고, 다음은 Multistix-SG, Combur-9-test RL의 순이었다. 요빌리루빈 및 요당검사에서는 두시험지봉이 비슷한 민감도를 나타내었다. 요중 백혈구검사에서는 요침사검사가, 요중 잠혈검사에서는 Combur-9-test RL이 가장 민감하였다.

• 대한수의학회지
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• 제39권5호
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• pp.951-960
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• 1999

We performed this study to evaluate the potential clinical marker of urinary trypsinogen-2 together with amylase, lipase and urinary amylase creatinine clearance ratio (ACCR) for the diagnosis of acute pancreatitis in dogs. In the experiment on daily changing patterns of amylase, lipase and ACCR measurements in experimentally induced pancreatitis dogs, compared to values measured in pre-induction state, significant difference was seen in amylase until 5th day of induction, and for lipase significant difference was found during the 7th day of observation period (p < 0.05). No significant difference was found in ACCR for the study period (p > 0.05). On SDS-PAGE analysis of urine from experimentally induced pancreatitis dog, The 26kd band was markedly increased compared with that of normal state and that band was confirmed trypsinogen-2 using substrate interaction and isoelectric focusing assay after being eluted. When assessing the appearance of 26kd band on urine SDS-PAGE 87.1% (range: 50~100%) of experimentally induced pancreatitis dogs showed positive results, whereas no corresponding band was seen in dog without pancreatic disorders. With this result, determination of urinary trypsinogen-2 assay was found to have a high diagnostic value with a 70% of sensitivity and 100% of specificity as a routine test for pancreatitis, although the detection of trypsinogen-2 in urine can be varied on the progression stage of pancreatitis at the initial visit to animal clinic. We therefore suggest that the promising results in this study be used for the development of dipstick test for detecting acute pancreatitis in the future research.

• Journal of Microbiology and Biotechnology
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• 제33권8호
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• pp.1091-1100
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• 2023

Human papillomavirus (HPV) types 16 and 18 are the major causes of cervical lesions and are associated with 71% of cervical cancer cases globally. However, public health infrastructures to support cervical cancer screening may be unavailable to women in low-resource areas. Therefore, sensitive, convenient, and cost-efficient diagnostic methods are required for the detection of HPV16/18. Here, we designed two novel methods, real-time ERA and ERA-LFD, based on enzymatic recombinase amplification (ERA) for quick point-of-care identification of the HPV E6/E7 genes. The entire detection process could be completed within 25 min at a constant low temperature (35-43℃), and the results of the combined methods could be present as the amplification curves or the bands presented on dipsticks and directly interpreted with the naked eye. The ERA assays evaluated using standard plasmids carrying the E6/E7 genes and clinical samples exhibited excellent specificity, as no cross-reaction with other common HPV types was observed. The detection limits of our ERA assays were 10⁰ and 10¹ copies/µl for HPV16 and 18 respectively, which were comparable to those of the real-time PCR assay. Assessment of the clinical performance of the ERA assays using 114 cervical tissue samples demonstrated that they are highly consistent with real-time PCR, the gold standard for HPV detection. This study demonstrated that ERA-based assays possess excellent sensitivity, specificity, and repeatability for HPV16 and HPV18 detection with great potential to become robust diagnostic tools in local hospitals and field studies.