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PubMed2006

[PCR-multiplex system for analysis of Y-chromosomal microsatellite polymorphisms DYS449, DYS456, DYS458, and DYS464].

DNA profiling--inclusive sex determination-- with microsatellite markers is currently a commonly used genetic method of studying humans. An efficient technique of producing the genetic data is amplification of multiple microsatellites in a single PCR reaction. Here we introduce a novel PCR-multiplex system for analysis of four polymorphic Y-STRs. Specifically, these are DYS449, DYS456, DYS458, and DYS464. These loci were chosen because of their reported high diversity in Euroamerican population (10), as well as their absence in the commercial analytical kits at the time of beginning of this study. Our objective was to design this PCR-multiplex for use of fragmentation analysis by electrophoresing samples on a capillary semi-automated genetic analyzer applying only one fluorescent dye. The PCR system we propose, may be notably used in fields such as forensic and human population genetics.

Chromosomes, Human, Y↗
PubMed2006

Differentiation of lymphocystis disease virus genotype by multiplex PCR.

Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease. The viruses have been divided into three genotypes (genotype I for LCDV-1, II for Japanese flounder isolates, and III for rockfish isolates) on the basis of major capsid protein (MCP) gene sequences. In this study, we developed a multiplex PCR primer set in order to distinguish these genotypes. We also analyzed the MCP gene of a new LCDV isolate from the sea bass (SB98Yosu). Comparison of sequence identities between SB98Yosu and eight Japanese flounder isolates, revealed identity of more than 90.1% at nucleotide level and 96.5% at deduced amino acid level, respectively. Phylogenetic analyses based on the MCP gene showed that SB98Yosu belongs to genotype II, along with Japanese flounder isolates. Multiplex PCR based on the MCP gene allowed us to identify these genotypes in a simple and rapid manner, even in a sample that contained two genotypes, in this case genotypes II and III.

Base Sequence↗
PubMed1991

Deletion analysis of Duchenne muscular dystrophy using cDNA probes and multiplex PCR.

The authors studied Duchenne muscular dystrophy (DMD) in 14 Yugoslav families with 16 diseased men. All the sixteen patients showed typical phenotypic features of DMD. The linkage analysis was done with five probes 754, C7, PERT87.15, pERT87.30, and pERTBir. A deletion with pERT87.15 (DX 164) in one family was found. Deletion analysis was mainly performed using cDNA probes and multiplex PCR analysis. Preferential deletions were in the hot spot deletion region covered by cDNA probe 8. Among 14 examined patients with cDNA hybridization deletions were found in 5 cases (5/14). For quick and prenatal analysis the authors used multiplex PCR. With this methods deletions were found in 4 cases (4/15). The percentage of the observed deletions cases was 50% (8/16) in Yugoslav population, which shows good correlation with other European and American populations studied. Prenatal diagnosis was done for two consultants with highly ambiguous carrier risk, and no deletions were found.

Child↗
PubMed2006

A successful strategy for Preimplantation Genetic Diagnosis of beta-thalassemia and simultaneous detection of Down's syndrome using multiplex fluorescent PCR.

OBJECTIVES: Preimplantation Genetic Diagnosis (PGD) is an alternative to prenatal diagnosis providing couples the chance to start a pregnancy with an unaffected fetus. The objective of the present study was to develop and apply quick, sensitive and accurate single cell PCR protocols for PGD of beta-thalassemia and Down's syndrome detection. MATERIAL AND METHOD: Two couples carrying beta-thalassemia codon41-42 mutation underwent routine IVF procedures. Embryo biopsy was performed on Day-3 post-fertilisation and single cell multiplex fluorescent PCR was employed for mutation analysis, contamination detection and diagnosis of trisomy 21 cases. RESULTS: Seventeen embryos were tested in two clinical PGD cycles. This resulted in the first birth following PGD for a single gene disorder in Thailand and South East Asia, confirmed by prenatal testing. Two embryos were shown to be affected by Down's syndrome. CONCLUSION: Successful strategy for PGD of beta-thalassemia and Down's syndrome detection using multiplex fluorescent PCR was introduced.

Adult↗
PubMed2005

[Application of the molecular test PCR multiplex for identification of Mycobacterium bovis BCG strains].

In our last paper (18) we described the problem of proper microbiological identification of BCG strains and how important is distinguishing vaccine strain from virulent strains of Mycobacterium tuberculosis complex. We have suggested the modern algorithm of BCG strains identification including mycolic acids profile by HPLC and 14C PZA resistance methods. These methods allowed us to made fast and accurate microbiological identification of side effects of BCG vaccine in the children. Identification of BCG by HPLC is possible within one working day compared with 3-4 weeks required for conventional methods. However both methods need very expensive instruments like HPLC and/or Bactec-460 Tb radiometric system. Presently we have evaluated molecular test based on the analyzis of the region RD1 encoding a 9.5-kb fragment. This fragment is deleted in all BCG substrains (6) and present in all human and bovine virulent strains. To evaluate this method for the rapid and specific detection of BCG, a large strain collection (32 strains) representating M. bovis BCG (vaccine strains and strains isolated from the children in case of adverse reactions after vaccination) M. bovis and M. tuberculosis from own collection was analyzed. RD1 was present in all 15 M. tuberculosis and M. bovis tested strains and deleted in 17 of 18 BCG strains. The multiplex PCR method was 100% sensitive and specific for the identification of BCG among strains of the Mycobacterium tuberculosis complex. Multiplex PCR can be used as a diagnostic test and has significant advantages over existing methods.

Animals↗
PubMed2006

[Species identification by multiplex amplifying mtDNA-HV I, HV II and cytb regions].

OBJECTIVE: To evaluate the method for species identification by multiplex amplification of mtDNA-HV I, HV II and cytb regions. METHODS: The bloodstains or muscular tissues of human and 16 kinds of animals were collected. DNA were quantified after extraction. Three pairs of primers were mixed in the same reaction and the PCR amplification products were analyzed by PAGE and silver staining. RESULTS: The presence of three bands (278 bp, 358 bp, 425 bp) indicated the samples were from human, while only one band indicated nonhuman origin. The position of the animal's only band is different from the band (358 bp) of human. CONCLUSION: The method of species identification by multiplex amplifying mtDNA-HV I, HV II and cytb regions is simple and sensitive. It is of use for forensic identification.

Animals↗
PubMed2006

One-tube multiplex PCR method for rapid identification of mycobacterium tuberculosis.

A rapid, inexpensive, simple, and accurate multiplex polymerase chain reaction (PCR) was developed in a single tube for identification of Mycobacterium tuberculosis. Assessment of sensitivity and specificity of simple PCR was performed with 116 strains of M. tuberculosis complex (MTC) and 144 strains of nontuberculous mycobacteria (NTM) compared with the biochemical method. Specific amplification of KS4, MTC-specific DNA fragment, was found in 98% (114/116) of MTC and not detected in 99% (143/144) of NTM. Amplification of the mtp40 gene revealed 95% sensitivity (100/105 strains of M. tuberculosis) and 77% specificity (not found in 119/155 mycobacterial strains). A multiplex PCR method based on the combination of KS4- and mtp40-derived primers was used for identification of M. tuberculosis. Crude DNA from slow growing mycobacteria with cream rough colonies that showed both 768-bp amplified product for KS4 and 396-bp for mtp40 was identified as M. tuberculosis whereas that from MTC gave only the 768-bp product.

Base Sequence↗
PubMed2006

[Establishment of miniCTTA multiplex set with fluorescence-labeled primers].

OBJECTIVE: To establish a miniCTTA multiplex set including short tandem repeat (STR) markers as CSF1PO, TH01, and TPOX and amelogenin gene, whose amplified fragments are shorter than 130 bp. METHODS: The length of the gene fragments amplified with fluorescence-labeled primers were analyzed by ABI3100 Genetic Analyzer for genotyping of 100 unrelated individuals, 10 genealogies and 30 highly degraded specimens. RESULTS: The genotypes derived form miniCTTA were consistent with the results by AmpFLSTR Identifiler kit. CONCLUSION: MiniCTTA multiplex set is useful for personal identification and paternity test, especially applicable for degraded DNA sample.

Amelogenin↗
PubMed2006

The M and N genes-based simplex and multiplex PCRs are better than the F or H gene-based simplex PCR for Peste-des-petits-ruminants virus.

Nucleocapsid (N), matrix (M) and hemagglutinin (H) genes-based simplex PCRs and an N and M genes-based multiplex PCR were developed for detection of Peste-des-petits-ruminants virus (PPRV). The M gene PCR was the most sensitive, followed by N, H and an already described fusion (F) gene PCRs, as they could detect the virus in samples with titers of 101, 102, 104and 105 TCID50/ml, respectively. The multiplex PCR was as sensitive as the M gene PCR, but it had the advantage of differentiating PPRV from Rinderpest virus (RPV).

Electrophoresis, Agar Gel↗
PubMed2006

[Detection of IgVH mutation status in patients with chronic lymphocytic leukemia by multiplex PCR].

IgVH mutation status is one of the most important independent prognostic factor of chronic lymphocytic leukemia (CLL). In order to evaluate IgVH mutation status in patients with CLL, IgVH mutation was detected by multiplex PCR in 9 CLL patients and purified PCR amplification products were directly sequenced, IgH somatic hypermutation and mutation site were analysed by IMGT/V-QUEST. The results showed that 5 patients had mutated IgVH, and IgVHs were IGHV3-11*03, IGHV3-9*01, IGHV3-23*01, IGHV4-59*01, IGHV4-34*02, respectively; whereas 4 others had unmutated IgVH, these IgVHs were IGHV3-53*01, IGHV3-23*03, IGHV3-33*05 and IGHV3-7*01. It is concluded that multiplex PCR is a rapid and easy method to detect IgVH mutation status, and it solves the limitations and pitfalls of routine PCR, and it is worth being extensively used both in clinic and scientific researches.

Adult↗
PubMed1991

[Mono-radiculopathy multiplex--multiple infarction of the cauda equina caused by intravascular lymphomatosis].

A 55-year-old man had felt numbness of the bilateral peroneal sides of legs for 6 months. Then hepatosplenomegaly, anemia, body weight loss and fever developed, and a diagnosis of malignant histiocytosis (MH) was made by revealing the presence of innumerable atypical histiocytes with hemophagocytosis in the bone marrow. Soon later, sensory disturbance of bilateral peroneal sides of legs (right side dominant) developed and aggravated with painful dysesthesia and weakness of the legs for the last 2 weeks before death. Electrophysiologically, sensory conduction velocity of the sural nerve was normal and somatosensory evoked potentials from tibial nerve were normal before P15 but were not evoked at all after the lumbar potential, suggesting lumbosacral radiculopathy. Autopsy showed multifocal ischemic lesions and secondary degeneration of the lumbosacral nerve roots associated with necrosis and fibrosis of the radicular vessels and intravascular infiltration of atypical mononuclear cells which were positive for B cell markers. The neurological manifestations and the distribution of ischemic lesions, which were similar to those of vasculopathic mononeuropathy multiplex, would deserve the name of "monoradiculopathy multiplex".

Cauda Equina↗
PubMed1991

Prevalence of autoantibodies in simplex and multiplex families of Brazilian insulin-dependent diabetic patients.

1. Forty-one simplex and 6 multiplex families of Brazilian IDDM patients were studied by the indirect immunofluorescence technique to determine the prevalence of the following autoantibodies: islet cells (ICA), islet cells which fix complement (ICA-CF), thyroid microsomes (TMA), thyroglobulin (TGA), and gastric-parietal cells (PCA). A total of 54 IDDM patients belonging to two family groups were analyzed. 2. A significantly higher frequency of ICA-CF and TMA was detected among the siblings from multiplex families than among those from simplex families (18.7% vs 1.7%). 3. A prospective study of 5 ICA-positive siblings was undertaken, and 2 who later became diabetic were found to be positive to both ICA and ICA-CF. 4. The prevalence of islet-cell antibodies in these 54 Brazilian IDDM patients and their unaffected first-degree relatives from genetically mixed groups suggests that the humoral autoimmune mechanisms of the disease are probably identical to those observed in other populations of different ethnic backgrounds.

Adolescent↗
PubMed1991

Application of a novel chemiluminescence-based DNA detection method to single-vector and multiplex DNA sequencing.

A chemiluminescent DNA detection method is described and its application shown for both single-vector and multiplex DNA sequencing using the standard dideoxy chain-termination process. This recently developed detection method, which utilizes the light emitted by an enzyme-catalyzed dioxetane reaction, is highly sensitive and affords significant advantages in safety and speed over the traditional radioactive labeling method. When adapted to a multiplex strategy, this chemiluminescent detection method constitutes a safe, simple and rapid method for increasing the throughput of DNA sequencing procedures.

Base Sequence↗
PubMed1990

[Sevoflurane anesthesia for a patient with arthrogryposis multiplex congenita].

We reported an anesthetic experience of a 6-year-old male patient with arthrogryposis multiplex congenita who underwent tonsillectomy and adenoidectomy. The induction and maintenance of anesthesia were smoothly performed with inhalation of 1-5% sevoflurane in nitrous oxide and oxygen. Emergence from anesthesia was also smooth. Halothane and succinylcholine chloride were not used to avoid possible hyperthermia. Anesthetic management of patients with arthrogryposis multiplex congenita was discussed citing literature references.

Adenoidectomy↗
PubMed1985

Thermal environmental monitoring of divers using a twelve channel multiplexing instrument.

A 12-channel temperature monitoring instrument designed for use by divers while immersed or within pressure chambers is described. Data are multiplexed at the diver and transmitted to the surface, bell, submarine, or chamber exterior through a two-core electrical cable. In the case of the immersed diver the cable may be incorporated into the diver's umbilical cable. Details of the method and special construction techniques used in the design of the diver-borne multiplexer are given. At the dive control station the 12 individual temperatures may be displayed in either digital or analogue format. The temperature data may also be stored on one track of a battery-powered magnetic tape recorder for subsequent off-line analysis.

Body Temperature↗
PubMed1984

[Steatocystoma multiplex].

A 50 years old male patient with numerous cysts over the scalp since the age of 48 is reported in whom the lesions appeared abruptly in two crops in a short period of time. Several biopsies were done showing most of them the typical histologic characteristics of steatocystoma multiplex. The authors remark in this patient the nonhereditary character of the condition, appearing late in life with an eruptive character and strict limitation over the scalp, features that are not often seen in steatocystoma multiplex. The authors pointed out that not all the cysts showed the typical histopathological picture of the disease. Therefore several biopsies must sometimes be done, before the diagnosis is established.

Epidermal Cyst↗
PubMedDate not supplied

[Case of "Friedreich's paramyoclonus multiplex". Clinical and neurophysiologic study].

The author reports the case of 13 years old girl, who since at least three years displays very frequent partial involuntary contractions of some arm's and leg's muscles, especially on the right but also on the left side. The girl was examined with EEG and contemporary registration of the myogramm of the muscles in question and with EMG. The DD of the myoclonic syndrome, and especially between Schultze myochimie, Kny's fibrillar myoclonus and Friedereich paramyoclonus multiplex has been discussed. The conclusion was that the case under discussion, is a case of paramyoclonus multiplex of Friedereich.

Adolescent↗