Ultrasensitive Nonradioactive Detection of PCR Reactions: An Overview

Tullis, Richard H.

doi:10.1007/978-1-4612-0257-8_10

Cited by 2 publications

(1 citation statement)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is possible, however, to adapt PCR assays to a variety of other detection methods (e.g. Tullis, 1994), in which the specificity of the PCR product is determined not by size but by the incorporation of either a primer-specific label such as a fluorescent dye (see Heller, 1994) or a specific ligand that will subsequently bind to a detection molecule (see Tu & Jablonski, 1994). These approaches usually require detection equipment as well as the additional cost per diagnostic PCR reaction associated with each the detection method, but they can offer two significant advantages.…”

Section: Modifications Of Pcr For Species Diagnosismentioning

confidence: 99%

A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species

Collins

Paskewitz

1996

Insect Molecular Biology

360

270

View full text Add to dashboard Cite

Cryptic species complexes are groups of closely related species that are difficult or impossible to distinguish by morphological traits. These complexes are known from a wide variety of arthropods and are common among the well-studied, medically-important insects. For example, many of the anopheline vectors of malaria parasites are members of cryptic species complexes. Complexes typically include both vector and non-vector species, and two or more member species are often found syrnpatrically. Until the late 195Os, only two such Anopheles complexes were known, the A. gambiaecomplex from Africa and the A. maculipennis complex from Europe. Today, dozens of Anopheles cryptic species complexes are recognized, and accumulating evidence suggests that most important malaria vectors are likely to be members of such complexes.A variety of methods have been developed for identifying the species of individual specimens from these complexes, although until recently only those based on species-specific allozymes and polytene chromosome inversions were widely used. The limitations inherent in these methods have been circumvented with DNA-based procedures, which are especially useful because both sexes and all developmental stages can be identified, and DNA can be recovered from samples stored by a wide variety of simple methods. Several DNA-based identification techniques have been developed, including hybridization assays based on species-specific repeat sequences, and diagnostic PCR fragments produced

show abstract

Section: Modifications Of Pcr For Species Diagnosismentioning

confidence: 99%

A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species

Collins

Paskewitz

1996

Insect Molecular Biology

360

270

View full text Add to dashboard Cite

show abstract

Molecular Detection of Diaporthe phaseolorum and Phomopsis longicolla from Soybean Seeds

Zhang¹,

Hartman²,

Curio-Penny³

et al. 1999

Phytopathology®

View full text Add to dashboard Cite

Species-specific detection of Diaporthe phaseolorum and Phomopsis longicolla from soybean seeds was accomplished using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and TaqMan chemistry. To use these detection systems, fungal DNA was released from soybean seed coats using an ultrasonic processor to break the cells. DNA fragment lengths ranged from 200 to 1,200 base pairs (bp), with the majority of fragments <500 bp. Based on DNA sequences of the internal transcribed spacer (ITS) regions of ribosomal DNA, three TaqMan primer/probe sets were designed. Primer/probe set PL-5 amplified a 96-bp fragment within the ITS1 region of P. longicolla, D. phaseolorum var. caulivora, D. phaseolorum var. meridionalis, and D. phaseolorum var. sojae. Set PL-3 amplified a 86-bp DNA fragment within the ITS2 region of P. longicolla. Set DPC-3 amplified a 151-bp DNA fragment within the ITS2 region of D. phaseolorum var. caulivora. TaqMan primer/probe sets were able to detect as little as 0.15 fg (four copies) of plasmid DNA. When using PCR-RFLP for Diaporthe and Phomopsis detection, the sensitivity was as low as 100 pg of pure DNA. Among 13 soybean seed lots from Italy and the United States, the total Diaporthe and Phomopsis detected using a traditional seed-plating technique ranged from 0 to 32%. P. longicolla was most prevalent, followed by D. phaseolorum var. sojae. D. phaseolorum var. caulivora, which only occurred in 0.5% of the Italian seed lots, was not detected in the U.S. seed lots. D. phaseolorum var. meridionalis was not detected in either the U.S. or Italian seed lots. Using TaqMan primer/probe set PL-3, the frequency of P. longicolla was 18% in seed lot I3, similar to the frequency obtained from PCR-RFLP and potato dextrose agar plating detection. The frequencies of D. phaseolorum and P. longicolla in each seed lot obtained by the different detection methods were comparable with respect to total infection and individual species detection. However, TaqMan detection provided the fastest results of all the methods tested.

show abstract

Ultrasensitive Nonradioactive Detection of PCR Reactions: An Overview

Cited by 2 publications

References 64 publications

A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species

A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species

Molecular Detection of Diaporthe phaseolorum and Phomopsis longicolla from Soybean Seeds

Contact Info

Product

Resources

About