Immunochromatographic Assay Kit Research Articles

Development of an on-site plum pox virus detection kit based on immunochromatography

Sharka disease, caused by plum pox virus (PPV), is the most serious viral disease of stone fruit trees. Among the eight known strains of the virus, PPV-D is the most important due to its recent global spread. Although enzyme-linked immunosorbent assay (ELISA) is the most common approach for diagnosing sharka, it involves time-consuming steps and requires expensive equipment and trained technicians. In this study, an on-site PPV detection kit based on immunochromatography was developed using polyclonal antibodies against the coat protein (CP) of a PPV-D isolate. The immunochromatographic (IC) assay kit was as sensitive as a commercial ELISA system for detecting Japanese PPV-D isolates. Moreover, it was easy to use (a one-step procedure), and results could be obtained on-site within 15 min without special laboratory equipment. The IC assay kit detected the virus from every aerial part of symptomatic Japanese apricot trees. In a detailed study of viral localization in leaves, the most suitable plant parts for use in the IC assay were symptomatic mesophyll tissues and the region from the petiole to the main vein. A positive reaction was also observed using the CP of other major (PPV-M and PPV-Rec) and minor (PPV-EA, PPV-W, and PPV-T) strains.

イムノクロマト診断キットによる温州萎縮ウイルスの周年検定

SDVにより引き起こされる温州萎縮病は収量や果実品質の低下を引き起こすことから問題となっている．カンキツのウイルス検定はこれまでELISA法が一般的に用いられてきたが，新梢を検体とすることから検定できる期間が1年のうち数週間と限定されており，かつ検定に時間を要することが問題となっていた．そこで，イムノクロマト診断キットにより簡単，迅速に，かつ周年可能なSDV検出方法を検討した．その結果，春の新梢と同様に，花については花弁，柱頭，子房の部位別に分離して検定したところ，いずれの部位を用いても検出可能であった．夏季から秋季にかけては，果皮のフラベド部分を用いると高率に検出が可能であり，特に日光の照射量が多い夏季は内成り果の果皮を用いると検出率が高かった．冬季においては，3ヶ月間常温貯蔵庫で貯蔵した果実の果皮からも検出可能であった．夏季採取の果実や長期貯蔵果実でも検出が可能であったが，検体によっては検出率が低下する事例が確認されたので検定する際はなるべく多くの果実を供試する必要がある．また,カンキツ苗木育成のための穂木のウイルス診断に際しては，冬季切り枝水挿し法による新梢や花蕾を用いても陽性反応が確認された．これらの結果から，SDVクロマトによる検定に新梢，花蕾，果皮，冬季の切り枝から発生させた新梢や花蕾を用いることでSDVの周年検出が可能となった．

Clinical evaluation of highly sensitive silver amplification immunochromatography systems for rapid diagnosis of influenza

In this study, the clinical usefulness of silver amplification immunochromatography (SAI) influenza virus detection kits, which employed a photographic development technology to increase the sensitivity of the conventional immunochromatographic assay was evaluated. Influenza A and B virus strains of nasopharyngeal aspirates obtained from influenza patients were tested at different dilutions on the SAI system and conventional immunochromatographic assay kit (ESPLINE Influenza A & B-N), and detection limits were calculated for comparison. The detection ability of the SAI system was 8 times higher for Influenza A viruses and 32–64 times higher for Influenza B viruses. Then 1118 respiratory specimens were obtained from patients who presented with influenza-like symptoms between 2009 and 2012. The sensitivities of the SAI system were 91.2% for type A and 94.4% for type B viruses and higher than those of the conventional kit. The SAI system also showed excellent specificities, 95.8% for type A and 98.0% for type B viruses, and was able to detect influenza viruses even within 6h after the disease onset with 90% sensitivity. In conclusion, the SAI system is useful for diagnosis of influenza from early stages of the illness.

Multicentre clinical study of the herpes simplex virus immunochromatographic assay kit for the diagnosis of herpetic epithelial keratitis

Background/aimsThe novel immunochromatographic assay (ICGA) kit was recently developed to diagnose herpes simplex virus (HSV) infection. This multicentre study aimed to evaluate the value of the ICGA kit for the...

Evaluation of a New Immunochromatographic Assay Kit for the Rapid Detection of Norovirus in Fecal Specimens

Rapid and accurate detection of norovirus is essential for the prevention and control of norovirus outbreaks. This study compared the effectiveness of a new immunochromatographic assay kit (SD BIOLINE Norovirus; Standard Diagnostics, Korea) and real-time reverse transcription-PCR (RT-PCR) for detecting norovirus in fecal specimens. Compared with real-time RT-PCR, the new assay had sensitivity, specificity, positive predictive value, and negative predictive value of 76.5% (52/68), 99.7% (342/343), 98.1% (52/53), and 95.5% (342/358), respectively. The sensitivity of the assay was 81.8% (18/22) for GII.3 and 75.7% (28/37) for GII.4. None of the 38 enteric virus-positive specimens (3 for astrovirus, 5 for enteric adenovirus, and 30 for rotavirus) tested positive in the cross-reactivity test performed by using this assay. The new immunochromatographic assay may be a useful screening tool for the rapid detection of norovirus in sporadic and outbreak cases; however, negative results may require confirmatory assays of greater sensitivity.

Validation of an immunochromatographic assay kit for the identification of the Mycobacterium tuberculosis complex

The performance of the immunochromatographic assay, SD BIOLINE TB Ag MPT64 RAPID®, was evaluated in Madagascar. Using mouse anti-MPT64 monoclonal antibodies for rapid discrimination between the Mycobacterium tuberculosis complex and nontuberculous mycobacteria, the kit was tested on mycobacteria and other pathogens using conventional methods as the gold standard. The results presented here indicate that this kit has excellent sensitivity (100%) and specificity (100%) compared to standard biochemical detection and can be easily used for the rapid identification of M. tuberculosis complex.

Rapid Detection of Infectious Bursal Disease Virus (IBDV) in Chickens by an Immunochromatographic Assay Kit

닭 전염성 F낭병 (IBD)은 닭에서 전염성이 강하고 발병으로 인하여 양계 산업에 막대한 경제적 피해를 입히는 닭의 바이러스성 전염병이다. 이 연구에서는 수 분 이내에 검사 시료로부터 닭 전염성 F낭병 바이러스를 검출할 수 있는 시판용 면역크로마토그래피법 검사 킷트를 이용하여 IBD 진단에 있어서의 유용성을 조사하였다. 사용한 면역크로마토그래피법 검사 킷트는 닭 전염성 F낭병 바이러스 VP2에 특이적인 단클론 항체를 이용하여 닭 전염성 F낭병 바이러스를 검출하도록 고안되었다. 바이러스 감염 역가를 알고 있는 IBDV를 사용하여 조사한 결과, IC 검사 킷트의 검출 한계는 <TEX>$10^{3.1}$</TEX> 내지 <TEX>$10^{3.9}$</TEX> <TEX>$EID_{50}$</TEX>/mL이었다. 이 검사 킷트는 닭의 다른 전염성 바이러스인 뉴캣슬병 바이러스, 닭 전염성 기관지염 바이러스, 조류인플루엔자 바이러스 및 전염성 후두기관염 바이러스에 대하여 비특이 반응을 나타내지 않았다. 고병원성의 IBDV를 실험적으로 감염시킨 후 3일 내지 4일에 폐사한 닭의 장기별로 조사한 결과, 모든 폐사 닭의 F낭, 장편도, 비장, 신장 시료들은 면역크로마토그래피법 검사 킷트에서 강한 양성반응을 나타내었다. 검사 시료 중 F낭 시료가 IC 검사 킷트에서 가장 강한 양성반응을 나타내었다. 폐사 닭의 간, 흉선, 선위의 경우, 각각 검사시료의 87.5%, 37.5% and 0%가 양성 반응을 나타내었다. 면역크로마토그래피법 검사 킷트에서 음성이었던 시료 중 흉선 시료 한 점을 제외한 모든 시료는 DAS-ELISA와 동일한 검사 결과를 나타내었으나, 검사 시료 중 흉선과 선위 일부에서 RT-PCR 검사에서 양성 반응을 나타내었다. 야외 IBD 발생 농장과 비발생 농장에서 수거한 폐사닭 231수의 조직을 면봉으로 도말하여 채취한 시료를 조사하여 RT-PCR법과 비교한 결과, 상대적 민감도와 특이도는 각각 100% (109/109) 및 97.5% (119/122)를 나타났으며, 두 검사 방법간 kappa value는 0.97이었다. 우리의 연구 결과는 IC 검사 킷트는 야외 양계 농장에서 폐사 닭을 대상으로 IBD를 진단하는 데 적용하기에 매우 유용하다는 것을 말해준다. An immunochromatograhy (IC) based infectious bursal disease virus (IBDV) detection kit, which employed two anti-IBDV VP2 monoclonal antibodies, was evaluated for rapid diagnosis of infectious bursal disease virus (IBD). The detection limit of the IC kit for IBDV was <TEX>$10^{3.1}$</TEX> to <TEX>$10^{3.9}$</TEX> <TEX>$EID_{50}$</TEX>/mL, indicating that the IC kit detected IBDV sensitively as same as double antigen capture ELISA but less than a RT-PCR assay. The IC kit did not detect other viral pathogens such as Newcastle disease virus, infectious bronchitis, avian influenza virus, and infectious larynotracheitis virus. When applied to tissue samples of experimental chickens died 3 or 4 days post infection after very virulent IBDV (strain Kr/D62) infection, the IC kit detected IBDV in all samples of the bursa of Fabricius, spleen, kidney, cecal tonsil and in 87.5%, 37.5% and 0% of liver, thymus and proventriculus samples. In particular, BF tissue samples showed stronger signal bands than other tissues. Positive signal was observed. All except for one thymus sample of samples having negative results by the IC kit showed the same result with DAS-ELISA but RT-PCR assay detected IBDV in some of IC kit negative samples of thymus and proventriculus. When swab samples from the bursa of Fabricius of dead chickens (n=231) on field farms were tested, the sensitivity and specificity of the IC assay relative to RT-PCR was 100% (109/109) and 97.5% (119/122), respectively and kappa value between both assay was 0.97. The kit can provide a useful aid for rapid detection of IBDV in chickens under field circumstances.

Comparison of Nine Different Qualitative HBsAg Assay Kits

Qualitative hepatitis B surface antigen (HBsAg) assay kits are still commonly used in Korea where hepatitis B virus (HBV) infection is endemic. The accurate determination of HBsAg plays a crucial role in the diagnosis and prevention of HBV infection, especially in endemic areas. The aim of this study was to compare the detection sensitivities of 9 qualitative HBsAg assay kits. Seven pooled sera with HBsAg concentration ranging from 0.14 IU/mL to 29.96 IU/mL were prepared. The HBsAg concentration of each pooled serum was determined by a quantitative HBsAg assay, Architect HBsAg (Abbott Laboratories, Ireland). The fully automated immunoassay kits included Elecsys HBsAg (Roche Diagnostics, Germany) and Immulite 2000 HBsAg (DPC, USA) and the rapid tests included 5 immunochromatographic assay (ICA) kits and 2 reverse passive hemagglutination assay (RPHA) kits. Elecsys HBsAg (Roche Diagnostics) showed positive result in pooled serum with HBsAg concentration of 0.14 IU/mL, but Immulite 2000 HBsAg (DPC) showed negative result in the same concentration. Although ICA kits showed variable results among different assay kits, all of them showed negative results in pooled sera with HBsAg concentration of < or = 1.89 IU/mL. Two RPHA kits showed negative results in pooled sera with HBsAg concentration of < or = 7.98 IU/mL. Although ICAs were more sensitive than RPHAs, they had variable sensitivities for HBsAg and were less sensitive than the automated immunoassay kits. Therefore, ICAs and RPHAs should be used with caution in the screening tests for HBsAg and their sensitivities need to be improved.

Evaluation of an Immunochromatographic Assay Kit for Rapid Identification of Mycobacterium tuberculosis Complex in Clinical Isolates

Evaluation of an Immunochromatographic Assay Kit for Rapid Identification of <i>Mycobacterium tuberculosis</i> Complex in Clinical Isolates

Simplified method for determining cadmium concentrations in rice foliage and soil by using a biosensor kit with immunochromatography

AbstractBACKGROUND: The behavior of cadmium in ecosystems needs to be monitored because of the human toxicity of this heavy metal. The need recently arose for a simple and quick on‐site test for trace levels of Cd in food and environmental samples. In response, an immunochromatographic assay kit for detecting Cd was manufactured by Kansai Electric Power Co. of Japan. This kit uses the antigen–antibody complex reaction between the Cd–EDTA complex and an anti‐Cd–EDTA antibody and shows the results in terms of the degree of color developed on a test paper. We previously reported the successful use of this kit to determine Cd concentrations in brown rice. Here, we applied the kit to the determination of Cd concentrations in rice foliage and soil.RESULTS: Cadmium in rice foliage was not extracted successfully by the method used for brown rice. However, it was successfully extracted by 0.1 mol L−1 HCl solution at a rice foliage:HCl ratio of 1:20, and coexisting metals were removed sufficiently by the column treatment. The Cd concentrations determined by immunochromatographic assay were well correlated with the values obtained by acid decomposition and inductively coupled plasma mass spectrometry. The 0.1 mol L−1 HCl‐extractable Cd concentration in soil was also determined successfully with the kit.CONCLUSION: Approximate Cd concentrations in rice plants and 0.1 mol L−1 HCl‐extractable Cd concentrations in soil can be monitored easily and quickly by this method at locations where facilities for acid digestion and precision analysis are not available. Copyright © 2009 Society of Chemical Industry

Evaluation of Immunochromatographic Assay for Serodiagnosis of Brucella canis

Canine brucellosis is a contagious disease with venereal and oral modes of transmission that produces late abortion in females, epididymides and prostates in males. Diagnosis is difficult because of unstable serum antibody titers that vary from individual to individual as well as between different methods used for their detection. The objective of this work was to evaluate the clinical utility of the immunochromatographic assay (ICA) for serodiagnosis of dogs suspected of having brucellosis, and results were compared with those obtained for hemoculture (HC) and the rapid screening agglutination with 2-mercaptoethanol (2-ME RSAT). The all experimentally infected dogs were positive in ICA, HC and 2-ME RSAT from 5 weeks, 7 weeks, and 3 weeks after infection, respectively. Also, among dogs selected from 10 different breed kennels occurred brucellosis, 24.8%, 39.5% and 39.1% of dogs tested were detected as positive with HC, 2-ME RSAT and ICA, respectively. The kappa value between 2-ME RSAT and ICA was 0.89. The results of this study showed that sensitivity and specificity of the ICA are comparable with those obtained using conventional serological and bacteriological test for brucellosis. In conclusion, the ICA kit provides a handy and accurate tool for the rapid serodiagnosis of canine brucellosis.

Evaluation of Duopath Legionella Kit for the Rapid Identification of Legionella Strains Isolated from Water Samples

Duopath Legionella (Merck KGaA, Darmstadt, Germany) is a rapid and simple immunochromatographic assay kit for the identification of Legionella species. We evaluated the precision of the kit in identifying 100 strains of Legionella and 35 strains of non-Legionella bacteria isolated from cooling tower and bath water samples. Consequently, of all the Legionella strains tested, 99 strains were judged to be Legionella, and only one strain (Legionella busanensis) was judged to be non-Legionella. All of the 35 non-Legionella strains were judged to be non-Legionella. We therefore conclude that Duopath Legionella is a useful method for the rapid identification of Legionella.

イムノクロマトグラフィーを用いたロタウイルスとアデノウイルスの迅速診断キットの検討

We investigated the usefulness of Rapidtesta Rota-Adeno (Daiichi Pure Chemicals Co., Ltd., Japan) for rapid detection of group A rotavirus and adenovirus simultaneously using immunochromatography with clinical samples. In investigation of the reaction of the kit to 5 strains of group A rotavirus, 13 strains of adenovirus and other intestinal viruses, specific lines were formed in red to group A rotavirus and blue to adenovirus. No cross reaction was observed with other intestinal virus. In measurement of detection limit, group A rotavirus (SA11) was detected at 10(4.4)TCID50/ml, serotype 3 adenovirus at 10(4.45)TCID50/ml. The detection limit of the kit was similar to other immunochromatographic assay or enzyme immnoassay kits and approximately 10 times higher than that of kits using latex agglutination test. In comparison with other testing kits in clinical samples, the concordance with other immunochromatographic assay was 99.2% (121/122) in group A rotavirus and 98.6% (138/140) in adenovirus. The rate of concordance with latex agglutination test kit was 94.5% (69/73) in group A rotavirus and 92.3% (84/91) in adenovirus. The kit had high rates of concordance with other immunochromatographic assay kits and higher virus detection rates than those of latex agglutination test kits. Rapidtesta Rota-Adeno is able to detect group A rotavirus and adenovirus simply and rapidly. In addition, the two kinds of virus can be easily differentiated by color difference in reaction lines, suggesting that the kit is useful in clinical diagnosis.

Evaluation of rapid immunochromatographic assay kit for HBsAg-screening using whole blood

A rapid immunochromatographic assay kit using whole blood to screen hepatitis B surface antigen was developed and evaluated by using sera from 240 patients. The reference diagnosis was based on the results obtained with GENEDIA Anti-HBs Rapid kit which is very similar to the above kit except for the use of serum. The test demonstrated a good corre- lation with the reference immunochromatographic assay kit, that is, the sensitivity and the specificity of the kit was 100%, respectively. The rapid test kit using whole blood should be more convenient and useful for the diagnosis of hepatitis B virus because the kit does not need machines and time to prepare serum. In addition, this kit is safe from inadvertent infec- tion during sample treatment because the blood is sterilized with hydrogen peroxide, elimi- nates the procedure required to prepare serum and reduces the possibility of exposure to in- fectious agents.