Applied DNA Sciences Inc (APDN)

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Apdn (b.v.i.) Inc. Patent Application – Methods for Genetic Analysis of Textiles Made of Gossypium Barbadense and Gossypium Hirsutum Cotton

2014 Oct 21 — By a News Reporter-Staff News Editor at Life Science Weekly -- According to news reporting originating from Washington, D.C., by NewsRx journalists, a patent application by the inventors LIANG, MingHwa Benjamin (East Setauket, NY); Shu Kin So, Stephane (Stony Brook, NY), filed on February 27, 2014, was made available online on October 9, 2014.

The assignee for this patent application is Apdn (b.v.i.) Inc.

Reporters obtained the following quote from the background information supplied by the inventors: “In general, there are two major types of cotton species being cultivated throughout the world, namely Gossypium barbadense and Gossypium hirsutum. These two species are from the genus Gossypium, which comprises at least 40 different cotton species. Gossypium barbadense (sea island cotton) and Gossypium hirsutum (upland cotton) are allotetraploids and are known as New World cotton or ‘American’ cotton. There are striking differences in the physical characteristics of the cotton fibers produced by G. barbadense compared to the cotton fibers produced by G. hirsutum. G. barbadense produces longer cotton fibers than most other cotton species and these fibers are usually called ‘extra long staple’ (ELS) fibers, while those of G. hirsutum are shorter and are called or defined as ‘upland’ fibers. Textiles made of ELS fibers are considered to be of higher quality compared to textiles made with shorter fibers, like those produced by G. hirsutum cotton plants. Thus, textiles produced from ELS cotton fibers are considered more valuable in the textile marketplace.

“There have been many studies trying to manipulate cotton genes for fiber quality improvement (U.S. Pat. Nos. 6,169,174; 7,060,874; and 6,995,256), enhanced pesticide toxin production (U.S. Pat. Nos. 6,686,149; 6,140,075; and 6,057,370), and herbicide resistance (U.S. Pat. No. 7,223,906). There have also been studies investigating the genetic polymorphism of various cotton species using PCR-based markers (J. Applied Sci. Res. 3(10)1156-1169, 2007). However, no success has been found in the categorization of cotton cultivars using genetic markers on mature cotton fibers mainly because of the lack efficient primers to amplify fragmented DNA in mature cotton fibers.

“Unfortunately, once cotton fibers are processed and made into yarn and/or fabrics most physical properties have been altered, and there is no reliable method to determine the origin or species of the fibers utilized to produce the yarn or textile(s). Forging clothing or producing knock-off textile items is a serious problem for the textile industry, costing manufacturers and retail stores millions and perhaps billions of dollars annually, in the United States alone. Being able to identify the species of cotton utilized in a textile item would not only be a way to authenticate an item as legitimate, but would also enable the detection of forged or counterfeit textile products.”

In addition to obtaining background information on this patent application, NewsRx editors also obtained the inventors’ summary information for this patent application: “Methods for authenticating articles of manufacture containing cotton fibers such as garments or other textile goods are described. Higher quality cotton textile goods are typically made from Gossypium barbadense known to have Extra Long Staple (ELS) cotton fibers. Counterfeit textile goods are often made with different, inferior cotton with shorter fibers, such as Gossypium hirsutum. When the cotton species in an original textile article is known, the detection of a different species of cotton in the textile articles subsequent to manufacture will indicate that a counterfeit article has been substituted. The methods provide a means for identifying the cotton species in a textile item from a sample of mature cotton fibers taken from the textile item, and for authenticating the textile items based on the identification of the cotton species. The authentication can be carried out at different points along the supply or commerce chain between manufacture and retail sale to identify the point in the chain at which counterfeit goods are introduced.

“In one embodiment, the method for identifying cotton species comprises collecting cotton fibers from a sample, extracting genomic DNA from the collected cotton fibers, and amplifying the DNA with PCR-based techniques, followed by analyzing the amplified product to distinguish between a first cotton species and a second cotton species. In the illustrative embodiment the first cotton species is G. barbadense and the second cotton species is G. hirsutum. In other embodiments, the first cotton species is selected from the group consisting of G. barbadense, G. hirsutum, G. arboreum or G. herbaceum. In still other embodiments, the second cotton species is selected from the group consisting of G. barbadense, G. hirsutum, G. arboreum or G. herbaceum.

“Further, in some embodiments the sample is a textile item while in other embodiments the sample is raw cotton material.

“In certain embodiments, the amplifying further comprises using at least one set of specific primers which targets sequence polymorphism between the first cotton species and the second cotton species.

“In some embodiments, the genomic DNA extracted from the cotton fibers originating from the textile in question comprises chloroplast DNA and in other embodiments it comprises nuclear DNA.

“In certain embodiments, the targeted sequences of the first cotton species and the second cotton species have different detectable physical characteristics.

“In some embodiments, the detectable physical characteristic of the targeted sequence in most embodiments is a difference in sequence length for the first cotton species compared to the second cotton species.

“In other embodiments, the detectable physical characteristic of the targeted sequence is a different sequence composition for the first and second cotton species.

“In most embodiments the analyzing of the amplified products comprises detecting the size of the amplified products by capillary electrophoresis. In other embodiments gel electrophoresis may be utilized instead of capillary electrophoresis.

“One embodiment of the method for verifying a textile article comprises collecting cotton fibers from a textile item, extracting genomic DNA from the cotton fibers collected, amplifying said DNA with PCR-based techniques, analyzing the amplified product to distinguish between a first cotton species and a second cotton species, identifying the cotton fibers from the textile item as belonging to the first cotton species or to the second cotton species, and determining from the identified cotton species if the textile article is authentic. In most embodiments for verifying a textile article, the first cotton species is G. barbadense and the second cotton species is G. hirsutum. In some embodiments for verifying a textile article, the amplifying further comprises using at least one set of specific primers which target sequence polymorphism between the first cotton species and the second cotton species. Also, the targeted sequences of the first cotton species and the second cotton species have different detectable physical characteristics. The detectable physical characteristic can be a difference in length of the targeted sequence when comparing the first and second cotton species to one another. The detectable physical characteristic can also be a slight variation in sequence composition.

“In one embodiment of a kit useful for carrying out the methods of the invention, the kit comprises a sample collection tube for placing mature cotton fibers obtained from the textile item, a DNA extraction solution, at least one set of specific primers specific for a target sequence found in a first cotton species and a second cotton species, as well as a PCR amplification buffer solution.

“In some embodiments the kit for determining the cotton species of a sample or item further comprises a PCR instrument.

“In yet other embodiments, the kit may further be found comprising an internal control for comparing the size of the amplified product.

“In certain embodiments, the kit comprises a capillary electrophoresis device.

“All patents and publications identified herein are incorporated herein by reference in their entirety.


BRIEF DESCRIPTION OF THE FIGURES

“FIG. 1 is a diagram showing the comparison of a genetic variable region of two different cotton.

“FIG. 2 is a flow chart of one embodiment of the methods for identifying the cotton species of mature cotton fibers.

“FIG. 3 is a flow chart of one embodiment of the methods for authenticating an article.

“FIG. 4 is a photograph of an electrophoresis gel of DNA extracted from mature cotton fibers of known cultivars.

“FIG. 5 is an electrogram of a capillary electrophoresis analysis of DNA extracted from cotton fabrics of known materials; Sample 1 is made of ELS cottons and Sample 2 is made of upland cottons.”

For more information, see this patent application: LIANG, MingHwa Benjamin; Shu Kin So, Stephane. Methods for Genetic Analysis of Textiles Made of Gossypium Barbadense and Gossypium Hirsutum Cotton. Filed February 27, 2014 and posted October 9, 2014. Patent URL: http://appft.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&u=%2Fnetahtml%2FPTO%2Fsearch-adv.html&r=3133&p=63&f=G&l=50&d=PG01&S1=20141002.PD.&OS=PD/20141002&RS=PD/20141002