scholarly journals Development of sensitive detection methods using chemiluminescent enzyme immunoassays.

1998 ◽  
Vol 42 (4) ◽  
pp. 281-285
Author(s):  
Yoshihiro Ashihara ◽  
Mitsuo Isomura
Keyword(s):  
Sensitive Detection ◽  
Detection Methods ◽  
Enzyme Immunoassays

scholarly journals Spread and Increase of Ratoon Stunting Disease of Sugarcane and Comparison of Disease Detection Methods

Plant Disease ◽  
1999 ◽  
Vol 83 (12) ◽  
pp. 1170-1175 ◽  
Author(s):  
J. W. Hoy ◽  
M. P. Grisham ◽  
K. E. Damann
Keyword(s):  
Yield Loss ◽  
Detection Method ◽  
Xylem Sap ◽  
Disease Spread ◽  
Detection Methods ◽  
Disease Detection ◽  
Correct Identification ◽  
Dot Blot ◽  
Ratoon Stunting Disease ◽  
Enzyme Immunoassays

The spread and increase of ratoon stunting disease (RSD) resulting from two mechanical harvests were compared in eight sugarcane cultivars at two locations. RSD spread and increase were detected in the ratoon crops grown after each harvest and varied among cultivars and locations. Disease spread and increase were greater in plants grown from stalks collected at the first harvest than in the first ratoon growth from the harvested field. RSD infection was determined using five disease detection methods: alkaline-induced metaxylem autofluorescence; microscopic examination of xylem sap; and dot blot, evaporative-binding, and tissue blot enzyme immunoassays. The tissue blot enzyme immunoassay was the most accurate RSD detection method. The dot blot and evaporative-binding enzyme immunoassays were the least sensitive for detection of RSD-infected stalks, and alkaline-induced metaxylem autofluorescence was least accurate for correct identification of noninfected stalks. The results indicate that disease spread and increase are variable even among cultivars susceptible to yield loss due to RSD, and the greatest threat of disease spread and increase occurs at planting.

Gold nanoparticles in the clinical laboratory: principles of preparation and applications

2012 ◽  
Vol 50 (2) ◽  
Author(s):  
Hassan M.E. Azzazy ◽  
Mai M.H. Mansour ◽  
Tamer M. Samir ◽  
Ricardo Franco
Keyword(s):  
Gold Nanoparticles ◽  
Clinical Laboratory ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Practical Applications ◽  
Molecular Assays ◽  
Detection Of Biomarkers

AbstractIn order to meet the challenges of effective healthcare, the clinical laboratory is constantly striving to improve testing sensitivity while reducing the required time and cost. Gold nanoparticles (AuNPs) are proposed as one of the most promising tools to meet such goals. They have unique optophysical properties which enable sensitive detection of biomarkers, and are easily amenable to modification for use in different assay formats including immunoassays and molecular assays. Additionally, their preparation is relatively simple and their detection methods are quite versatile. AuNPs are showing substantial promise for effective practical applications and commercial utilization is already underway. This article covers the principles of preparation of AuNPs and their use for development of different diagnostic platforms.

Highly sensitive and rapid detection of citrus Huanglongbing pathogen (Candidatus Liberibacter asiaticus) using Cas12a-based methods

2021 ◽  
Author(s):  
Matthew Wheatley ◽  
Yong-Ping Duan ◽  
Yinong Yang
Keyword(s):  
Early Detection ◽  
Nucleic Acids ◽  
Detection Sensitivity ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Candidatus Liberibacter Asiaticus ◽  
Fluorescent Reporter ◽  
Candidatus Liberibacter ◽  
Citrus Huanglongbing ◽  
Liberibacter Asiaticus

Citrus Huanglongbing (HLB) or greening is one of the most devastating diseases of citrus worldwide. Sensitive detection of its causal agent, Candidatus Liberibacter asiaticus (CLas), is critical for early diagnosis and successful management of HLB. However, current nucleic acid-based detection methods are often insufficient for the early detection of CLas from asymptomatic tissue, and unsuitable for high-throughput and field-deployable diagnosis of HLB. Here we report the development of the Cas12a-based DETECTR (DNA endonuclease-targeted CRISPR trans reporter) assay for highly specific and sensitive detection of CLas nucleic acids from infected samples. The DETECTR assay, which targets the five-copy nrdB gene specific to CLas, couples isothermal amplification with Cas12a trans-cleavage of fluorescent reporter oligos and enables detection of CLas nucleic acids at the attomolar level. The DETECTR assay was capable of specifically detecting the presence of CLas across different infected citrus, periwinkle and psyllid samples, and shown to be compatible with lateral flow assay technology for potential field-deployable diagnosis. The improvements in detection sensitivity and flexibility of the DETECTR technology position the assay as a potentially suitable tool for early detection of CLas in infected regions.

A novel NIR fluorescent probe for highly sensitive detection of HNO and its application in bioimaging

2021 ◽  
Author(s):  
shenwei he ◽  
Jianming Zhu ◽  
peiyao xie ◽  
Jianfei Liu ◽  
Di Zhang ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Reactive Nitrogen Species ◽  
Physiological Process ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Reactive Nitrogen ◽  
Nitrogen Species ◽  
Efficient Detection ◽  
Highly Sensitive ◽  
Highly Sensitive Detection

Nitroxyl (HNO) as an important reactive nitrogen species (RNS), which plays an important role in multiple physiological process. Therefore, it is urgent to exploit reliable and efficient detection methods for...

scholarly journals Highly Sensitive Detection of PCV2 Based on Tyramide Signals and GNPL Amplification

Molecules ◽  
2019 ◽  
Vol 24 (23) ◽  
pp. 4364
Author(s):  
Shouping Zhang ◽  
Bin Hu ◽  
Xiaojing Xia ◽  
Yanzhao Xu ◽  
Bolin Hang ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Secondary Infection ◽  
Porcine Circovirus Type ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Porcine Circovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Binding Ability ◽  
Strong Binding ◽  
Dual Signal Amplification

The frequent emergence of secondary infection and immunosuppression after porcine circovirus type 2 (PCV2) infection highlights the need to develop sensitive detection methods. A dual-signal amplification enzyme-linked immunosorbent assay (ELISA) based on a microplate coated with gold nanoparticle layers (GNPL) and tyramide signal amplification (TSA) was established. Results confirmed that the microplates coated with GNPL have a strong binding ability to the antibody without affecting the biological activity of the antibody. The microplates coated with GNPL have strong binding ability to the antibody, and the amplification of the tyramide signal is combined to further improve the sensitivity of PCV2. The PCV2 antibody does not crossreact with other viruses, demonstrating that the method has good specificity. A dual-signal amplification strategy is developed using microplates modified with GNPL and TSA to sensitively detect PCV2.

scholarly journals Highly Sensitive GMO Detection Using Real-Time PCR with a Large Amount of DNA Template: Single-Laboratory Validation

2018 ◽  
Vol 101 (2) ◽  
pp. 507-514 ◽  
Author(s):  
Junichi Mano ◽  
Shuko Hatano ◽  
Yasuaki Nagatomi ◽  
Satoshi Futo ◽  
Reona Takabatake ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Sensitive Detection ◽  
Detection Methods ◽  
35S Promoter ◽  
Processed Foods ◽  
Gmo Detection ◽  
Nos Terminator ◽  
Highly Sensitive ◽  
Dna Template

Abstract Current genetically modified organism (GMO) detection methods allow for sensitive detection. However, a further increase in sensitivity will enable more efficient testing for large grain samples and reliable testing for processed foods. In this study, we investigated real-time PCR-based GMO detection methods using a large amount of DNA template. We selected target sequences that are commonly introduced into many kinds of GM crops, i.e., 35S promoter and nopaline synthase (NOS) terminator. This makes the newly developed method applicable to a wide range of GMOs, including some unauthorized ones. The estimated LOD of the new method was 0.005% of GM maize events; to the best of our knowledge, this method is the most sensitive among the GM maize detection methods for which the LOD was evaluated in terms of GMO content. A 10-fold increase in the DNA amount as compared with the amount used under common testing conditions gave an approximately 10-fold reduction in the LOD without PCR inhibition. Our method is applicable to various analytical samples, including processed foods. The use of other primers and fluorescence probes would permit highly sensitive detection of various recombinant DNA sequences besides the 35S promoter and NOS terminator.

scholarly journals A Lateral Flow Strip Based on a Truncated Aptamer-Complementary Strand for Detection of Type-B Aflatoxins in Nuts and Dried Figs

Toxins ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 136 ◽  
Author(s):  
Zhilei Zhao ◽  
He Wang ◽  
Wenlei Zhai ◽  
Xiaoyuan Feng ◽  
Xia Fan ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Cdna Probe ◽  
Test Strip ◽  
Lateral Flow ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Complementary Strand ◽  
Type B ◽  
Experimental Conditions ◽  
Lateral Flow Strip

Type-B aflatoxins (AFB1 and AFB2) frequently contaminate food, especially nuts and fried figs, and seriously threaten human health; hence, it is necessary for the newly rapid and sensitive detection methods to prevent the consumption of potentially contaminated food. Here, a lateral flow aptasensor for the detection of type-B aflatoxins was developed. It is based on the use of fluorescent dye Cy5 as a label for the aptamer, and on the competition between type-B aflatoxins and the complementary DNA of the aptamer. This is the first time that the complementary strand of the aptamer has been used as the test line (T-line) to detect type-B aflatoxins. In addition, the truncated aptamer was used to improve the affinity with type-B aflatoxins in our study. Therefore, the lengths of aptamer and cDNA probe were optimized as key parameters for higher sensitivity. In addition, binding buffer and organic solvent were investigated. The results showed that the best pair for achieving improved sensitivity and accuracy in detecting AFB1 was formed by a shorter aptamer (32 bases) coupled with the probe complementary to the AFB1 binding region of the aptamer. Under the optimal experimental conditions, the test strip showed an excellent linear relationship in the range from 0.2 to 20 ng/mL with a limit of detection of 0.16 ng/mL. This aptamer-based strip was successfully applied to the determination of type-B aflatoxins in spiked and commercial peanuts, almonds, and dried figs, and the recoveries of the spiked samples were from 93.3%−112.0%. The aptamer-complementary strand-based lateral flow test strip is a potential alternative tool for the rapid and sensitive detection of type-B aflatoxins in nuts and dried figs. It is of help for monitoring aflatoxins to avoid the consumption of unsafe food.

scholarly journals Serological and Molecular Assays for Rapid and Sensitive Detection of Iris yellow spot virus Infection of Bulb and Seed Onion Crops

Plant Disease ◽  
2008 ◽  
Vol 92 (4) ◽  
pp. 588-594 ◽  
Author(s):  
H. R. Pappu ◽  
I. M. Rosales ◽  
K. L. Druffel
Keyword(s):  
Real Time ◽  
Rapid Detection ◽  
The United States ◽  
Iris Yellow Spot Virus ◽  
Yellow Spot ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Spot Virus

Iris yellow spot virus (IYSV) has spread rapidly in the United States and has become an important economic constraint to the production of both bulb and seed onion crops. Symptoms caused by IYSV may be confused with those caused by other fungal and bacterial pathogens and virus-specific, reliable, sensitive, and rapid detection methods would improve the diagnosis. Antiserum was produced to Escherichia coli-expressed nucleocapsid protein of IYSV and an indirect format of the enzyme-linked immunosorbent assay (ELISA) was developed. IYSV could be detected in onion tissue at dilutions of up to 1:1,000. An IYSV-specific primer pair was designed and used in a real-time reverse-transcription polymerase chain reaction (RT-PCR) assay for the rapid detection of IYSV. Compared with standard RT-PCR, real-time RT-PCR was more rapid and sensitive. A commercially available RNA extraction kit and a total nucleic acid extraction method were compared for the quality of the templates obtained for use in real-time RT-PCR and there was no difference in limits of detection. Availability of ELISA- and PCR-based rapid and sensitive detection methods would facilitate accurate virus diagnosis and aid in better understanding of the epidemiology of the disease and in development of management strategies.

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