Rapid detection of foot-and-mouth disease virus using reverse transcription recombinase polymerase amplification combined with a lateral flow dipstick.

Foot-and-mouth disease caused by foot-and-mouth disease virus (FMDV) is one of the most highly contagious diseases of domestic animals, and leads to enormous economic loss. Currently there are two main prevention and control strategies for the disease: eradication of the infected animals in FMDV free countries, and vaccination of the susceptible animals in countries with endemic FMDV infection. Early discovery and diagnosis of the source of infection is therefore integral to the containment of FMDV. In this study, a two-step reverse transcription recombinase polymerase amplification assay combined with lateral flow detection (RPA-LFD) was developed to detect FMDV. With incubation at 38 °C, a region of the 2B gene on the FMDV genome was successfully amplified within 20 min using specific primers and a probe. The amplified RPA product can be visualized on a lateral flow dipstick. The RPA-LFD assay was highly sensitive, detecting down to 10 copies of plasmid DNA. There was no cross-reactivity with other pathogens causing vesicular lesions. In addition, 143 clinical samples were used to compare RPA-LFD with real-time PCR, with 98.6% concordance between the assays. Therefore, the developed RPA-LFD assay provides a rapid, simple, highly promising approach to be used as point-of-care diagnostics in the field.

[Correlation of Gemin 3 SNP in the microRNA Biosynthesis Pathway with Risk and Prognosis of Patients with Non-Hodgkin's Lymphoma].

OBJECTIVE: To evaluate the correlation of single nucleotide polymorphisms (SNP) of Gemin 3(rs197412) in the miRNA biosynthesis with NHL cancer risk and overall prognosis. METHODS: miR-SNP were genotyped using PCR-ligase detection reaction(LAR, LCR) in NHL group of 230 non-Hodgkin lymphoma (NHL) patients and in control group of 120 healthy persons. The survival curves were drawn using the Kaplan-Meier method, and comparisons between the curves were made using the log-rank test. Multivariate survival analysis was performed by using a Cox proportional hazards model. RESULTS: The rs197412 genotype distribution difference was not statistically significant, in NHL and control group; the survival time of patients carrying the rs197412 TT genotype was significantly longer than that of the patients carrying the CC+CT genotype (P=0.007). In addition, rs197412 was independent from the survival of NHL patients by multivariate analysis (RR: 2.138,95% CI: 1.303-3.508, P<0.01). CONCLUSION: The single nucleotide polymorphisms of Gemin 3 (rs197412) in the miRNA processing are not related with NHL risk, but that may affect NHL survival.

Primary giant lymphoma of the right thigh: A case report and brief review of the literature.

Primary soft tissue non-Hodgkin lymphoma (NHL) of the extremities is very rare. The clinical features of NHL mimic those of other soft tissue tumors, particularly sarcoma; however, they should be differentiated, as the treatment and prognosis are completely different. In this study, the case of a 68-year-old female with a giant mass, movement disorder, numbness and painful sensations in the right thigh is presented. Magnetic resonance (MR) imaging revealed a huge circle-shaped mass. Fine needle aspiration cytology (FNAC) of the tumor demonstrated neoplastic small, round cells. The tentative diagnosis was of a mesenchymal sarcoma. The right thigh was amputated. On histological examination of the amputated extremity, the diagnosis was found to be large B cell lymphoma. Primary soft tissue NHL of the extremities is a systemic malignant disease and is sensitive to chemo-therapy and radiotherapy. The histological diagnosis should be identified as far as possible before the tumor is widely excised.

Replacement of H1 linker histone during bovine somatic cell nuclear transfer.

Linker histone variants are involved in regulation of chromosome organization and gene transcription; several subtypes are expressed in the maturing oocyte and developing embryo. In Xenopus and mice, the transition between linker histone variants occurred following nuclear transfer, and apparently contributed to donor nuclear reprogramming. To determine whether such linker histone replacement occurred after bovine nuclear transfer, red fluorescent protein (RFP) tagged H1e (somatic linker histone H1e) donor cells and Venus tagged H1foo eggs were created, enucleated eggs were injected with donor cells, and embryos were created by fusion. Using fluorescence microscopy, release of H1e in the donor nucleus, acquisition of H1foo by donor chromosomes, and the H1foo-to-H1e transition were observed in live cells. Linker histone replacement occurred more slowly in bovine than murine embryos. Low levels of diffuse red fluorescence (H1e) in the donor nucleus were detected 5 h after fusion, at which time green fluorescence (H1foo) had incorporated into donor chromosomes. However, complete replacement did not occur until 8 h after fusion. We concluded that the linker histone transition was sufficiently conserved among species, which provided further evidence regarding its important role in nuclear reprogramming.

Matrix metalloproteinase-2 promoter and tissue inhibitor of metalloproteinase-2 gene polymorphisms in non-Hodgkin's lymphoma.

Single nucleotide polymorphisms in the promoter regions of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMP) genes are associated with an adverse outcome in some cancers. We examined three polymorphisms: -1306C/T and -735C/T in MMP-2 and -418G/C in the TIMP-2 gene, using chain reaction restriction fragment length polymorphism typing analysis in 575 patients with non-Hodgkin's lymphoma (NHL). We examined the possible correlations between the three polymorphisms (MMP-2 (-1306C/T and -735C/T) and TIMP-2 gene (-418G/C)) and the clinical significance and survival rate in patients with NHL. The incidence of the CT, TT+CT genotypes and T allele of -735C/T was significantly higher in stage III and IV patients compared to stage I and II patients. In cases with bone marrow infiltration, the TT genotypes of the -1306C/T gene were significantly less frequent compared to CC genotypes. The CT, TT and CT+TT genotypes and T allele in patients exhibiting the -1306C/T polymorphism were significantly less frequent in patients with a large tumor size compared to a smaller tumor. The TT genotypes of the -735C/T polymorphism were more common in patients with a large tumor compared to those with a smaller tumor. The frequency of the -1306C/-735T haplotype in patients with a smaller tumor size was significantly higher compared to patients with a large tumor. The -1306T/-735C and -1306C/-735C haplotypes were significantly less frequent in patients with B-symptoms compared to those without. Interestingly, patients with the -735CT genotype exhibited a lower rate of survival. Our results demonstrate that certain MMP-2 and TIMP-2 gene polymorphisms potentially effect the progression or assessment of prognosis for NHL. This research warrants further, larger scale studies.