Amplification of a Variable Number of Tandem Repeats (VNTR) Locus (pMCT118) by the Polymerase Chain Reaction (PCR) and Its Application to Forensic Science

1990 ◽  
Vol 35 (5) ◽  
pp. 12944J ◽  
Author(s):  
Kentaro Kasai ◽  
Yusuke Nakamura ◽  
Ray White
Keyword(s):  
Polymerase Chain Reaction ◽  
Forensic Science ◽  
Tandem Repeats ◽  
Variable Number ◽  
Chain Reaction ◽  
Vntr Locus ◽  
Polymerase Chain

Tuberculosis in swine co-infected withMycobacterium aviumsubsp.hominissuisandMycobacterium bovisin a cluster from Argentina

2014 ◽  
Vol 143 (5) ◽  
pp. 966-974 ◽  
Author(s):  
S. BARANDIARAN ◽  
A. M. PÉREZ ◽  
A. K. GIOFFRÉ ◽  
M. MARTÍNEZ VIVOT ◽  
A. A. CATALDI ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Null Hypothesis ◽  
Tandem Repeats ◽  
Spatial Clustering ◽  
Variable Number ◽  
Molecular Tools ◽  
Chain Reaction ◽  
Analysis Techniques ◽  
Swine Population ◽  
Polymerase Chain

SUMMARYIn Argentina little is known about the epidemiology of tuberculosis (TB) infection in swine. We characterized the epidemiological dynamics ofMycobacterium aviumcomplex (MAC) infection in a swine population of Argentina using molecular tools and spatial analysis techniques. Isolates (n = 196) obtained from TB-like lesions (n = 200) were characterized by polymerase chain reaction. The isolates were positive to eitherM. bovis(IS6110) (n = 160) orM. avium(IS1245) (n= 16) while the remaining 20 (10·2%) isolates were positive to bothM. bovisandM. avium. The detection of both bacteria together suggests co-infection at the animal level. In addition, MAC-positive isolates (n = 36) were classified asM. aviumsubsp.avium(MAA) (n = 30) andM. aviumsubsp.hominissuis(MAH) (n = 6), which resulted in five genotypes when they were typed using mycobacterial interspersed repetitive unit, variable number of tandem repeats (MIRU-VNTR). One significant (P = 0·017) spatial clustering of genotypes was detected, in which the proportion of MAH isolates was larger than expected under the null hypothesis of even distribution of genotypes. These results show that in Argentina the proportion of TB cases in pigs caused byM. aviumis larger than that reported in earlier studies. The proportion ofM. bovis–MAC co-infections was also higher than in previous reports. These results provide valuable information on the epidemiology of MAC infection in swine in Argentina.

scholarly journals Amplification by the polymerase chain reaction of hypervariable regions of the human genome for evaluation of chimerism after bone marrow transplantation

Blood ◽  
1991 ◽  
Vol 77 (7) ◽  
pp. 1607-1615 ◽  
Author(s):  
L Ugozzoli ◽  
P Yam ◽  
LD Petz ◽  
GB Ferrara ◽  
RE Champlin ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Marrow Transplantation ◽  
Tandem Repeats ◽  
Limit Of Detection ◽  
Minor Component ◽  
Restriction Enzymes ◽  
Mixed Chimerism ◽  
Chain Reaction ◽  
Oligonucleotide Hybridization ◽  
Polymerase Chain

We combined the polymerase chain reaction (PCR) with oligonucleotide hybridization as a novel and sensitive technique to evaluate posttransplant chimerism. Specific oligonucleotides for hybridization were synthesized homologous to tandemly repetitive core sequences of regions with a variable number of tandem repeats (VNTRs). Polymorphisms at such loci result from allelic differences in the number of repeats. Primers flanking the repeat region of each of the corresponding VNTRs were used for amplification. Recipient and donor pretransplant DNA and recipient posttransplant DNA were amplified. The resultant fragments were analyzed after gel electrophoresis either by hybridization in-gel or after Southern transfer. To confirm our findings, we also performed standard assays of restriction fragment length polymorphisms (RFLPs). Evaluation of 13 selected cases indicated mixed chimerism (4), complete chimerism (5), recurrence of leukemia (2), and endogenous repopulation of hematopoiesis (2) after marrow transplantation. Sensitivity of the method was determined by mixing various proportions of recipient and donor DNA; the limit of detection of the minor component in a mixture was 0.1%. PCR data correlated with RFLP data in all cases except two in which PCR proved more sensitive than RFLP. PCR amplification of VNTRs combined with oligonucleotide hybridization is a novel technique for documenting posttransplant chimerism and has advantages over RFLP analysis: high sensitivity, use of small amounts of DNA (250 ng), ease of preparation of DNA, elimination of need for restriction enzymes, and the ability to complete studies in 2 days.

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