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Topic:Polymerase Chain Reaction

Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA sequences, allowing for detailed genetic analysis in horses. This method enables the detection and quantification of genetic material, facilitating research in areas such as genetic disorders, infectious diseases, and population genetics in equine species. PCR applications in horses include identifying pathogens, verifying parentage, and studying genetic variations. The technique's sensitivity and specificity make it a valuable tool in equine veterinary diagnostics and research. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and advancements of PCR in equine science.
Isolation of virulent Rhodococcus equi from native Japanese horses.
Comparative immunology, microbiology and infectious diseases    March 15, 2001   Volume 24, Issue 2 123-133 doi: 10.1016/s0147-9571(00)00022-9
Takai S, Ogawa K, Fukunaga N, Sasaki Y, Kakuda T, Tsubaki S, Anzai T.R. equi was isolated from soil samples obtained from the environment of seven native Japanese horse breeds (Hokkaido, Kiso, Noma, Misaki, Tokara, Miyako and Yonaguni) and from fecal samples collected from three native horse breeds (Hokkaido, Kiso and Misaki). Virulent R. equi at various levels (ranging from 0.5 to 12.9%) was isolated from the feces or soil environment of Hokkaido, Kiso and Misaki horses. Isolates were investigated both for the presence of 15- to 17-kDa antigens (virulence-associated protein antigens; VapA) by colony blotting, using the monoclonal antibody 10G5, and the gene of...
Evaluation of the specificity of five oligoprobes for identification of cyathostomin species from horses.
International journal for parasitology    March 10, 2001   Volume 31, Issue 2 197-204 doi: 10.1016/s0020-7519(00)00161-2
Hodgkinson JE, Love S, Lichtenfels JR, Palfreman S, Ramsey YH, Matthews JB.Here, we report evaluation of five oligoprobes designed from intergenic spacer (IGS) region sequences for identification of cyathostomin species. Oligoprobes were designed for identification of Cylicocyclus ashworthi, Cylicocyclus nassatus, Cylicostephanus longibursatus, Cylicostephanus goldi and a fifth probe designed to identify all members of this tribe. PCR amplification of IGS DNA from 16 cyathostomin species allowed sequence comparison and identification of four putative species-specific probes. Southern blotting of amplified products from 16 species showed that all probes were species-s...
Rapid PCR detection of Salmonella in horse faecal samples.
Veterinary microbiology    March 7, 2001   Volume 79, Issue 1 63-74 doi: 10.1016/s0378-1135(00)00340-0
Amavisit P, Browning GF, Lightfoot D, Church S, Anderson GA, Whithear KG, Markham PF.A rapid polymerase chain reaction (PCR) assay was developed for detecting Salmonella in faeces of horses and assessed on samples from horses admitted to a veterinary hospital. Direct detection was achieved by amplification of part of ompC after extraction of DNA from faeces using a spin column method to reduce the amount of inhibitory substances in samples. An internal positive control was included to detect false negative results. While the sensitivity of the PCR assay was less than culture when assessed on faeces inoculated with Salmonella, its sensitivity on faecal samples obtained from hor...
A random amplified polymorphic DNA polymerase chain reaction technique that differentiates between Neospora species.
The Journal of parasitology    February 24, 2001   Volume 86, Issue 6 1366-1368 doi: 10.1645/0022-3395(2000)086[1366:ARAPDP]2.0.CO;2
Spencer JA, Witherow AK, Blagburn BL.Neospora caninum is a recently described coccidial parasite that was first isolated from a dog in 1988 and has subsequently been shown to infect a wide range of mammals. Neospora hughesi, a new species of this genus, has recently been isolated from the spinal cord of horses showing clinical signs of equine protozoal myeloencephalitis. The random amplified polymorphic DNA polymerase chain reaction technique is capable of differentiating between N. caninum and N. hughesi.
Molecular cloning and sequencing of equine cDNA encoding serum amyloid A (SAA).
Veterinary immunology and immunopathology    January 4, 2001   Volume 77, Issue 3-4 321-327 doi: 10.1016/s0165-2427(00)00239-7
Ma Z, Mizukoshi T, Khatlani TS, Okuda M, Onishi T.The serum amyloid A (SAA) protein is a characteristic and sensitive acute phase reactant in all vertebrates investigated. We molecularly cloned the equine cDNA encoding SAA from the liver of a healthy horse by polymerase chain reaction (PCR). The cloned cDNA is 480 bases in length, and contains an open reading frame (ORF) of 387 nucleotides encoding a precursor SAA protein of 128 amino acids. The precursor of horse SAA seems to have an 18-residue signal peptide and differs from the reported amino acid sequences of the horse SAA by substitution of valine at residue 81. It shows high homology wi...
Fatal nonneurological EHV-1 infection in a yearling filly.
Veterinary pathology    December 6, 2000   Volume 37, Issue 6 672-676 doi: 10.1354/vp.37-6-672
Del Piero F, Wilkins PA, Timoney PJ, Kadushin J, Vogelbacker H, Lee JW, Berkowitz SJ, La Perle KM.A case of fatal nonneurological equine herpesvirus 1 (EHV-1) infection in a yearling filly is described. Gross lesions included extensive pulmonary edema, prominent laryngeal lymphoid follicles, and congestion and edema of the dorsal third ventricle choroid plexus. Histologically, there was vasculitis, hemorrhage, and edema in the lungs and dorsal third ventricle choroid plexus as well as mild intestinal crypt necrosis with occasional intranuclear inclusion bodies. The perivascular and vascular inflammatory infiltrates were comprised mainly of T lymphocytes and macrophages. EHV-1 antigen was i...
Control of strangles outbreaks by isolation of guttural pouch carriers identified using PCR and culture of Streptococcus equi.
Equine veterinary journal    November 28, 2000   Volume 32, Issue 6 515-526 doi: 10.2746/042516400777584721
Newton JR, Verheyen K, Talbot NC, Timoney JF, Wood JL, Lakhani KH, Chanter N.Previous use of repeated nasopharyngeal swabbing and culture of Streptococcus equi showed that healthy carriers developed in more than 50% of 'strangles' outbreaks. The guttural pouches were the only detectable site of S. equi colonisation on endoscopic examination of horses during one of these outbreaks and S. equi was sometimes not detected by culture of nasopharyngeal swabs from carriers for up to 2 or 3 months before nasal shedding resumed sporadically. A more sensitive way of detecting S. equi on swabs from established guttural pouch carriers was therefore required. Conveniently selected ...
Clinical application of a polymerase chain reaction assay in the diagnosis of pneumonia caused by Rhodococcus equi in a horse.
Journal of the American Veterinary Medical Association    November 4, 2000   Volume 217, Issue 9 1348-1350 doi: 10.2460/javma.2000.217.1348
Vivrette SL, Sellon DC, Gibbons DS.Diagnosis of pneumonia caused by Rhodococcus equi can be made more rapidly by use of a polymerase chain reaction (PCR) assay than by use of conventional bacteriologic culture techniques. Use of a PCR assay aids in the differentiation between virulent and avirulent strains of R equi, and the assay may be used to identify R equi in feces and soil of breeding farms.
Prevalence of equine herpesvirus type 1 latency detected by polymerase chain reaction.
Archives of virology    October 24, 2000   Volume 145, Issue 9 1773-1787 doi: 10.1007/s007050070055
Carvalho R, Oliveira AM, Souza AM, Passos LM, Martins AS.In this study, an improved polymerase chain reaction (PCR) was used for detection of DNA of latent EHV-1 strains from several sources. Three pairs of oligonucleotide primers spanning fragments of 333 bp, 226 bp and 268 bp of the thymidine kinase (tk) gene, and one primer pair spanning 225 bp of the glycoprotein C (gC) gene were used in specific amplifications. Primers for EHV-4 PCR were also designed. Restriction digests with TaqI confirmed the identity of tk PCR fragments from EHV-1. The sensitivity to detect PCR products was further improved by visualisation in silver-stained acrylamide gels...
Demonstration of equine herpesvirus-1 gene expression in the placental trophoblasts of naturally aborted equine fetuses.
Journal of comparative pathology    October 18, 2000   Volume 123, Issue 2-3 119-125 doi: 10.1053/jcpa.2000.0401
Mukaiya R, Kimura T, Ochiai K, Wada R, Umemura T.Equine herpesvirus-1 (EHV-1) infection was demonstrated in the lung tissue of seven aborted fetuses by immunohistochemical labelling and polymerase chain reaction. The placentas of the fetuses were also examined by non-isotopic in-situ hybridization for the EHV-1 glycoprotein B (gB) gene. Positive hybridization signals were observed in the cytoplasm of trophoblasts, especially in microcotyledons, of all seven placentas, and in villous epithelium of the allantochorion of six placentas. Despite the presence of EHV-1 RNA, EHV-1 antigens were not detected in placentas by immunohistochemical examin...
The beta-tubulin gene of Babesia and Theileria parasites is an informative marker for species discrimination.
International journal for parasitology    October 12, 2000   Volume 30, Issue 11 1181-1185 doi: 10.1016/s0020-7519(00)00105-3
Cacciò S, Cammà C, Onuma M, Severini C.A fragment of the beta-tubulin gene was polymerase chain reaction (PCR) amplified from genomic DNAs of Babesia bovis, Babesia bigemina, Babesia divergens, Babesia major, Babesia caballi, Babesia equi, Babesia microti, Theileria annulata and Theileria sergenti. Single amplification products were obtained for each of these species, but the size of the amplicons varied from 310 to 460 bp. Sequence analysis revealed that this variation is due to the presence of a single intron, which ranged from 20 to 170 bp. The extensive genetic variability at the beta-tubulin locus has been exploited to develop...
Genotyping of Bacteroides fragilis isolates from stool specimens by arbitrarily-primed-PCR.
Diagnostic microbiology and infectious disease    September 7, 2000   Volume 37, Issue 4 225-229 doi: 10.1016/s0732-8893(00)00150-4
Sarma PN, Tang YJ, Prindiville TP, Osborne PD, Jang S, Silva J, Cohen SH.In order to determine genetic relatedness of Bacteroides fragilis isolates from different clinical sources, arbitrarily primed polymerase chain reaction (PCR) (AP-PCR) was used to compare 17 strains isolated from patients with inflammatory bowel disease (IBD) and 20 strains isolated from foals with diarrhea. Three reference ATCC strains were also analyzed. Eighteen unique types were identified with a 22-mer arbitrary primer (ERIC-2) among the 20 patient isolates. Types 1 (enterotoxigenic) and 9 (nonenterotoxigenic), were each found in the stools of two patients. All other isolates showed a dis...
Experimental and natural borna disease virus infections: presence of viral RNA in cells of the peripheral blood.
Veterinary microbiology    September 6, 2000   Volume 76, Issue 3 229-244 doi: 10.1016/s0378-1135(00)00242-x
Vahlenkamp TW, Enbergs HK, Müller H.Cells of the peripheral blood of experimentally and naturally borna disease virus (BDV)-infected animals and of human psychiatric patients and healthy individuals were analyzed for the presence of viral RNA using a BDV-p40-specific nested reverse transcription-polymerase chain reaction (RT-PCR). The assay proved to be highly sensitive as 10 RNA molecules were reproducibly amplified. BDV RNA was detected in blood cells of experimentally infected immunocompetent mice and rats. Mice were persistently infected without showing clinical signs of borna disease (BD), whereas the rats suffered from acu...
Characterization of Escherichia coli isolated from adult horses with and without enteritis.
The veterinary quarterly    August 22, 2000   Volume 22, Issue 3 162-166 doi: 10.1080/01652176.2000.9695048
van Duijkeren E, van Asten AJ, Gaastra W.In the present study E. coli strains isolated from the faeces of ten horses with diarrhoea and 14 horses without diarrhoea were characterized. All horses were culture negative for Salmonella species. Nine colonies of E. coli from each faecal sample were picked at random and a DNA fingerprint was made by means of a polymerase chain reaction (PCR) using Enterobacterial Repetitive Intergenic Consensus (ERIC) primers. The number of E. coli genotypes did not differ significantly between horses with and without diarrhoea. In addition, all E. coli strains with different DNA fingerprints were tested b...
Determination of the cDNA sequence and mRNA expression of interleukin-1 receptor antagonist in horses.
American journal of veterinary research    August 22, 2000   Volume 61, Issue 8 920-924 doi: 10.2460/ajvr.2000.61.920
Dhar AK, Thompson MS, Paradis MR, Alcivar-Warren A.To determine the complementary DNA (cDNA) sequence of interleukin-1 receptor antagonist (IL-1ra) in horses and compare messenger RNA (mRNA) expression of IL-1ra among horses of various breeds. Methods: Blood samples from neonatal and adult horses examined for a variety of diseases. Methods: A polymerase chain reaction procedure was used to amplify a 220 base pair (bp) portion of the genomic DNA. The upstream and downstream regions of the cDNA sequence were determined by means of 5' and 3' rapid amplification of cDNA ends (RACE) procedures. Northern blot hybridization was used to examine steady...
Comparison of Northern blot hybridization and a reverse transcriptase-polymerase chain reaction technique for measurement of mRNA expression of metalloproteinases and matrix components in articular cartilage and synovial membrane from horses with osteoarthritis.
American journal of veterinary research    August 22, 2000   Volume 61, Issue 8 900-905 doi: 10.2460/ajvr.2000.61.900
Fehr JE, Trotter GW, Oxford JT, Hart DA.To determine relative amounts of mRNA expression of aggrecan, type-II collagen, matrix metalloproteinase (MMP) 1, and MMP3 in articular cartilage and synovial membrane samples from healthy equine joints and joints with osteoarthritis (OA) and to compare results of Northern blot hybridization with results of a reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Methods: Articular cartilage samples from 8 pairs of joints (1 with OA and 1 healthy) from 6 horses and synovial membrane samples from 6 pairs of joints from 5 horses. Methods: RNA was extracted from samples by use of a modif...
Development of a differential multiplex PCR assay for equine herpesvirus 1 and 4 as a diagnostic tool.
Journal of veterinary medicine. B, Infectious diseases and veterinary public health    July 20, 2000   Volume 47, Issue 5 351-359 doi: 10.1046/j.1439-0450.2000.00361.x
Carvalho R, Passos LM, Martins AS.In this study, a multiplex polymerase chain reaction (PCR) procedure was developed for differentiation of strains and field isolates of equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4). Specific oli-gonucleotide primers were combined to amplify the thymidine kinase (TK) gene region of EHV-1 and EHV-4, which would yield fragments of different lengths for each virus in the same amplification reaction. The specificity of the largest PCR amplicon for EHV-4 was confirmed by restriction digestion with HindIII. The multiplex PCR proved to be a fast and sensitive method for typing EHV-1 and EHV-4 ...
Detection of Leptospira spp. in the aqueous humor of horses with naturally acquired recurrent uveitis.
Journal of clinical microbiology    July 6, 2000   Volume 38, Issue 7 2731-2733 doi: 10.1128/JCM.38.7.2731-2733.2000
Faber NA, Crawford M, LeFebvre RB, Buyukmihci NC, Madigan JE, Willits NH.Leptospiral organisms have long been presumed to be associated with the presence of equine recurrent uveitis. This project was undertaken to determine the presence of Leptospira spp. in the aqueous humor of horses with uveitis to determine if there was an association with inflammation. Thirty horses were determined to have recurrent uveitis based on clinical evaluation or history. Sixteen horses were judged clinically and historically to be free of uveitis and were used as controls. Aqueous humor samples were cultured and evaluated by PCR for the presence of Leptospira DNA. Serum was collected...
Use of repetitive sequence-based polymerase chain reaction for molecular epidemiologic analysis of Streptococcus equi subspecies equi.
American journal of veterinary research    June 13, 2000   Volume 61, Issue 6 699-705 doi: 10.2460/ajvr.2000.61.699
Al-Ghamdi GM, Kapur V, Ames TR, Timoney JF, Love DN, Mellencamp MA.To determine whether repetitive sequence-based polymerase chain reaction (rep-PCR) could be used to differentiate Streptococcus equi isolates, to examine S equi isolates from throughout the world, and to determine whether a horse had > 1 subtype of S equi during an outbreak of disease. Methods: An initial group of 32 S equi isolates, 63 S equi isolates from various geographic areas, and 17 S equi isolates obtained during outbreaks of disease. Methods: An aliquot of S equi genomic DNA was amplified, using enterobacterial repetitive intergenic consensus primers. Gel electrophoresis was perfor...
Ehrlichia equi infection of horses from Minnesota and Wisconsin: detection of seroconversion and acute disease investigation.
Journal of veterinary internal medicine    June 1, 2000   Volume 14, Issue 3 252-257 doi: 10.1892/0891-6640(2000)014<0252:eiohfm>2.3.co;2
Bullock PM, Ames TR, Robinson RA, Greig B, Mellencamp MA, Dumler JS.Equine granulocytic ehrlichiosis (EGE) is caused by infection with Ehrlichia equi. EGE has been reported primarily in northern California, where E equi is transmitted by the tick Ixodes pacificus. Reports of EGE and the emergence of human granulocytic ehrlichia in Minnesota prompted a seroprevalence study of E equi in horses of Minnesota and Wisconsin. Tick (Ixodes scapularis) endemic areas of Minnesota and Wisconsin were compared to nonendemic regions of Minnesota. Indirect fluorescent antibody was used to detect the presence of serum antibodies to E equi. Serum samples from healthy horses, 3...
Primary nucleotide structure of predominant and alternate splice forms of equine insulin-like growth factor I and their gene expression patterns in tissues.
American journal of veterinary research    May 3, 2000   Volume 60, Issue 10 1234-1241 
Nixon AJ, Brower-Toland BD, Sandell LJ.To isolate, clone, and determine primary nucleotide sequence of equine insulin-like growth factor I (IGF-I) and to examine IGF-I gene expression in tissues and cartilage from horses. Methods: Horses of various ages. Methods: Total RNA was isolated from tissues and purified. Complementary DNA (cDNA) was derived by reverse transcription and polymerase chain reaction (PCR) amplification and subcloned to plasmid vectors for sequencing and comparison with other species. Total RNA from various tissues was probed with radiolabeled cDNA or complimentary RNA constructs by use of northern blotting, tube...
[Demonstration of Chlamydia from an equine abortion].
DTW. Deutsche tierarztliche Wochenschrift    April 1, 2000   Volume 107, Issue 2 49-52 
Henning K, Sachse K, Sting R.The isolation and identification of a chlamydial agent from an equine fetus is reported. The fetus was aborted by a mare with respiratory disease and fever in the 9th month of pregnancy. The serum of the mare was investigated by the compliment fixation test. Specific antibodies were detected for chlamydial antigen in a titer of > 1:40 and for equine herpes virus 1 antigen in a titer of 1:32. Pathological lesions were not found in the organs of the fetus. Chlamydiae were detected in the placenta by ELISA and subsequently isolated by cell culture. Using PCR technique the agent was identified ...
Detection of heterogeneous genotypes among Australian strains of Taylorella equigenitalis.
Australian veterinary journal    March 29, 2000   Volume 78, Issue 1 56-57 doi: 10.1111/j.1751-0813.2000.tb10362.x
Matsuda M, Kagawa S, Sakamoto Y, Miyajima M, Barton M, Moore JE.No abstract available
The genetic structure of Spanish Celtic horse breeds inferred from microsatellite data.
Animal genetics    February 26, 2000   Volume 31, Issue 1 39-48 doi: 10.1046/j.1365-2052.2000.00591.x
Cañon J, Checa ML, Carleos C, Vega-Pla JL, Vallejo M, Dunner S.Partition of the genetic variability, genetic structure and relationships among seven Spanish Celtic horse breeds were studied using PCR amplification of 13 microsatellites on 481 random individuals. In addition, 60 thoroughbred horses were included. The average observed heterozygosity and the mean number of alleles were higher for the Atlantic horse breeds than for the Balearic Islands breeds. Only eight percentage of the total genetic variability could be attributed to differences among breeds (mean FST approximately 0.08; P < 0.01). Atlantic breeds clearly form a separate cluster from th...
Determination of intraspecies variations of the V2 region of the 16S rRNA gene of Streptococcus equi subsp. zooepidemicus.
Research in veterinary science    February 24, 2000   Volume 68, Issue 1 33-39 doi: 10.1053/rvsc.1999.0332
Abdulmawjood A, Lämmler CH.The 16S rRNA gene of 39 S. equi subsp. zooepidemicus strains and two S. equi subsp. equi strains was amplified by polymerase chain reaction and subsequently digested with the restriction enzyme Hinc II. A restriction profile with two fragments with sizes of 1250 bp and 200 bp could be observed for both S. equi subsp. equi strains and for 30 of the 39 S. equi subsp. zooepidemicus strains indicating a sequence variation within the V2 region of the 16S rRNA gene of the remaining nine S. equi subsp. zooepidemicus isolates. A segment of the 16S rRNA gene including the hypervariable V2 region of 11 ...
Development of a PCR test for rapid diagnosis of contagious equine metritis.
The Journal of veterinary medical science    January 29, 2000   Volume 61, Issue 12 1287-1292 doi: 10.1292/jvms.61.1287
Anzai T, Eguchi M, Sekizaki T, Kamada M, Yamamoto K, Okuda T.In order to establish a rapid diagnostic method for contagious equine metritis (CEM), we developed and evaluated a polymerase chain reaction (PCR) test. Species-specific PCR primer sets were derived from the DNA sequence of a cloned DNA fragment of Taylorella equigenitalis that did not hybridize with the genome of a taxomonically related species, Oligella urethralis. Single step PCR with primer set P1-N2 and two-step semi-nested PCR with primer sets P1-N2 and P2-N2 detected as low as 100 and 10 CFU of the bacteria, respectively. Single-step PCR detected T. equigenitalis from genital swabs of e...
Experimental infection of ponies with Borrelia burgdorferi by exposure to Ixodid ticks.
Veterinary pathology    January 22, 2000   Volume 37, Issue 1 68-76 doi: 10.1354/vp.37-1-68
Chang YF, Novosol V, McDonough SP, Chang CF, Jacobson RH, Divers T, Quimby FW, Shin S, Lein DH.Seven specific-pathogen-free (SPF) ponies, 1-5 years old, were exposed to Borrelia burgdorferi-infected adult ticks while being treated with dexamethasone over 5 consecutive days. One SPF pony (pony No. 178) was first exposed to laboratory-reared nymphs without B. burgdorferi infection and 3 weeks later was exposed to B. burgdorferi-infected adult ticks with concurrent dexamethasone treatment for 5 consecutive days. Four uninfected ponies treated with dexamethasone, exposed to laboratory-reared ticks without B. burgdorferi infection served as uninfected controls. Clinical signs, bacteriologic ...
Detection of trypanosome DNA in serologically positive but aparasitaemic horses suspected of dourine in Ethiopia.
The Tokai journal of experimental and clinical medicine    January 6, 2000   Volume 23, Issue 6 303-308 
Clausen PH, Gebreselassie G, Abditcho S, Mehlitz D, Staak C.A field study of horses was conducted in the province of Bale, Ethiopian highlands. A rapid questionnaire analysis indicated that dourine, known as "Dirressa", is a major health problem of equines in this area. A total of 121 horses suspected of dourine were examined by use of clinical, parasitological, serological and DNA based techniques. Incoordination of hindlegs (76%), swelling of external genitalia (48.8%) and emaciation (39.7%) were the most common clinical signs observed. Using the haematocrit centrifugation technique (HCT), no trypanosomes were detected in blood, genital washes or tis...
Human granulocytic ehrlichiosis agent infection in a pony vaccinated with a Borrelia burgdorferi recombinant OspA vaccine and challenged by exposure to naturally infected ticks.
Clinical and diagnostic laboratory immunology    January 5, 2000   Volume 7, Issue 1 68-71 doi: 10.1128/CDLI.7.1.68-71.2000
Chang YF, McDonough SP, Chang CF, Shin KS, Yen W, Divers T.A pony was vaccinated with recombinant OspA vaccine (rOspA) and then exposed 3 months later to Borrelia burgdorferi-infected ticks (Ixodes scapularis) collected in Westchester County, N.Y. At 2 weeks after tick exposure, the pony developed a high fever (105 degrees F). Buffy coat smears showed that 20% of neutrophils contained ehrlichial inclusion bodies (morulae). Flunixin Meglumine (1 g daily) was given for 2 days, and the body temperature returned to normal. PCR for ehrlichial DNA was performed on blood samples for 10 consecutive days beginning when the pony was first febrile. This pony was...
Quantitative real-time PCR for equine cytokine mRNA in nondecalcified bone tissue embedded in methyl methacrylate.
Calcified tissue international    December 14, 1999   Volume 65, Issue 5 378-383 doi: 10.1007/s002239900717
Leutenegger CM, von Rechenberg B, Huder JB, Zlinsky K, Mislin C, Akens MK, Auer J, Lutz H.Specific amplification and quantitation of nucleic acid sequences by the polymerase chain reaction (PCR) has been extensively used for the detection of viral infection and gene expression. Although successful amplification of DNA and RNA sequences extracted from paraffin embedded tissue have been described, there are presently no reports available regarding RNA analysis from bone and calcified tissues embedded in hydrophobic acrylic resin. Here we describe a general method for quantitation of specific mRNA sequences extracted from undecalcified bone sections, fixed in paraformaldehyde, and emb...
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