-
[show abstract]
[hide abstract]
ABSTRACT: The H6 subtype of avian influenza virus (AIV) infection occurs frequently in wild and domestic birds. AIV antigen detection is preferred for controlling AIV as birds are infected before they produce antibodies. The purpose of this study was to develop an early diagnostic method for AIV detection. Six monoclonal antibodies (mAbs) developed from a field H6N1 AIV strain were tested for their ability to bind to viruses. The two that showed the greatest binding ability to AIVs were used for antigen detection. An antigen-capture enzyme-linked immunosorbent assay (ELISA) to detect H6 AIVs was developed using these mAbs. One mAb was coated onto an ELISA plate as the capture antibody. The other mAb was used as the detector antibody after labeling with horseradish peroxidase. The antigen-capture ELISA detected H6N1 AIVs but not H5 AIVs, human H1N1, H3N2 influenza or other viruses. This antigen-capture ELISA could be used to specifically detect H6N1 AIV.
Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi 12/2011; 45(3):243-7. · 0.99 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Pangolin scales are encountered in traditional East Asian medicines (TEAM) and the ever increasing demand for these scales has escalated the decline in the numbers of these mammals. The identification of protected pangolin species is necessary to enforce international and national legislation as well as assist with conservation measures. There is limited morphological feature on a pangolin scale thus requiring DNA analysis as a means of identification. We report on the isolation of DNA from pangolin scales and a strategy for obtaining the full length of the mitochondrial D-loop, being 1159 bp. Primer sets creating five overlapping amplicons were designed to amplify sections of this mitochondrial DNA locus. DNA from the blood stain of nineteen Formosan pangolins (Manis pentadactyla pentadactyla) along with 145 scale samples that were suspected to have come from pangolins, was amplified and sequenced; leading to a total of 91 D-loop sequences being obtained. The 19 Formosan pangolin sequences produced 5 haplotypes and 72 of the 145 seized scales provided useable sequence classified as a further 38 haplotypes. The D-loop sequences from those scales suspected to be from a pangolin had a higher similarity to any of the 19 samples taken from M. p. pentadactyla compared to a D-loop sequence from Manis tetradactyla (the only pangolin D-loop sequence in GenBank, NC_004027). These 43 haplotypes were used to establish a local database for the D-loop sequence of pangolins and add to the data of Manis sp. held on GenBank. The PCR amplification strategy development in this study could be used in forensic DNA identification of scales suspected to be from protected pangolin species.
Forensic science international. Genetics 08/2011; 5(4):303-7. · 2.42 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We report on a novel and rapid strategy for the simultaneous identification of both avian species and gender by analyzing a section of the CHD gene. The CHD gene is carried by the avian sex determining chromosomes where a female bird carries both a W and Z chromosome but a cock bird carries two copies of the Z chromosome. Two primer pairs, CHD1F/CHD1R and P2/P8, were used to amplify a part of the CHD gene from 144 samples corresponding to 58 avian species. For all species tested, two fragments were observed at least in one amplification for female samples. All tested species produced species specific size fragments allowing both sex determination and species identification using these primer pairs. However, special care is still warranted as so few samples have been characterised. This novel strategy for avian species and gender identification using the CHD gene was developed for a number of applications from ecology to forensic science.
Molecular and Cellular Probes 09/2009; 24(1):27-31. · 2.08 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We report a DNA-based test that can be applied to any avian species so that the amplicon can be used in species identification. The need for the test arose from the requirement to enforce the Wildlife Conservation Act in Taiwan where over 150 avian species are protected. It is difficult to enforce the law if no gross morphology is present and hence there is a requirement to develop a DNA test. This study uses a novel strategy for avian species identification by the cytochrome b gene where a series of primer pairs producing amplicons of decreasing size was designed. The test is designed to produce the largest possible amplicon based upon the quality of the DNA in the sample. A total of 331 avian samples were tested representing 40 species. Sequencing of the amplicons revealed limited intraspecies variation and that no DNA sequence was shared by samples from two different avian species. The closest genetic distance among the 40 species was 0.059 which was between Lonchura punctulata and Estrilda melpoda based upon data from the smallest amplicon. A DNA databank including 138 sequence types from 331 samples tested, representing 40 different species, was constructed in this study. A blind test was used to determine the value for this system for forensic applications that successfully identified the species.
Electrophoresis 07/2008; 29(11):2413-8. · 3.30 Impact Factor
-
Lih-Chiann Wang,
Chu-Hsiang Pan,
Lucia Liu Severinghaus,
Lu-Yuan Liu,
Chi-Tsong Chen,
Chang-En Pu,
Dean Huang,
Jihn-Tsair Lir,
Shih-Chien Chin,
Ming-Chu Cheng,
Shu-Hwae Lee,
Ching-Ho Wang
[show abstract]
[hide abstract]
ABSTRACT: Newcastle disease (ND) and avian influenza (AI) are two of the most important zoonotic viral diseases of birds throughout the world. These two viruses often have a great impact upon the poultry industry. Both viruses are associated with transmission from wild to domestic birds, and often display similar signs that need to be differentiated. A rapid surveillance among wild and domestic birds is important for early disease detection and intervention, and is the basis for what measures should be taken. The surveillance, thus, should be able to differentiate the diseases and provide a detailed analysis of the virus strains. Here, we described a fast, simultaneous and inexpensive approach to the detection of Newcastle disease virus (NDV) and avian influenza virus (AIV) using oligonucleotide microarrays. The NDV pathotypes and the AIV haemagglutinin subtypes H5 and H7 were determined at the same time. Different probes on a microarray targeting the same gene were implemented in order to encompass the diversified virus strains or provide multiple confirmations of the genotype. This ensures good sensitivity and specificity among divergent viruses. Twenty-four virus isolates and twenty-four various combinations of the viruses were tested in this study. All viruses were successfully detected and typed. The hybridization results on microarrays were clearly identified with the naked eyes, with no further imaging equipment needed. The results demonstrate that the detection and typing of multiple viruses can be performed simultaneously and easily using oligonucleotide microarrays. The proposed method may provide potential for rapid surveillance and differential diagnosis of these two important zoonoses in both wild and domestic birds.
Veterinary Microbiology 04/2008; 127(3-4):217-26. · 3.33 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Identifying the sex of a bird is important to ensure successful breeding strategies and effective conservation programs. Sex may be identified from the intron size of the CHD1 gene located on the avian sex chromosomes Z and W. However, because of the great nucleotide diversity across different avian species, no given intron is in widespread use without ambiguous results. Complicated modifications of the reaction condition are required to suit different species. Two CHD1 introns were used with a unified reaction condition in this study to simplify the procedure. Consequently, genders of 73 avian species covering 19 families were successfully identified based on this two-intron approach. This means the ability to sex a wider range of avian species using a simplified procedure, greatly assisting in population management at zoos. Zoo Biol 26:425-431, 2007. (c) 2007 Wiley-Liss, Inc.
Zoo Biology 10/2007; 26(5):425-31. · 0.84 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Sex determination of birds is important to ensure successful breeding strategies, especially for endangered species. Most birds are vulnerable to stress during handling, however, so obtaining a sufficient amount of genomic DNA (gDNA) while causing the least amount of harm is a critical issue. Avian gender can be determined based on different CHD1 gene intron sizes in W and Z sex chromosomes. We have compared various specimen sources and have found that the rachis segment of a feather is a good DNA source for determining sex. This indicates that plucking the whole feather is not necessary; a cut feather including the rachis is a superior method because it decreases stress on the examined birds and is accomplished easily. Zoo Biol. 0:1–5, 2006. © 2006 Wiley-Liss, Inc.
Zoo Biology 02/2006; 25(4):279 - 283. · 0.84 Impact Factor
-
Lih Chiann Wang,
Lucia Liu Severinghaus,
Chi Tsong Chen,
Lu Yuan Liu,
Chu Hsiang Pan,
Dean Huang,
Hsiao Yuan Lee,
Jihn Tsair Lir,
Shih Chien Chin,
Chang En Pu,
Ching Ho Wang
[show abstract]
[hide abstract]
ABSTRACT: Molecular sexing of the diversified avian family Strigidae is difficult. Sex identification using the intron length difference between W and Z chromosomal CHD1 genes, as visualized by agarose gel electrophoreses, often produces ambiguous results. Here we describe a simple method for sexing a variety of Strigidae species using oligonucleotide microarrays, on which several sex-specific probes operated complementarily or in concert. The sex of 8 owl species was identified clearly on the microarrays through sequence recognition. This sequence-directed method can be easily applied to a wider range of Strigidae species.
The Journal of heredity 99(2):187-92. · 2.05 Impact Factor
