Valutazione Della Carica Virale Del Citomegalovirus Umano (Hcmv)

Bergallo, Massimiliano; Merlino, C.; Tarallo, Sonia; Forgnone, F.; Sinesi, Franca; Baro, Sangeeta; penna, A. La; Ponzi, A. Negro; Cavallo, Rossana

doi:10.4081/mm.2005.3568

Cited by 6 publications

(6 citation statements)

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“…In the previous examples proteinase K was added to HRP in equimass quantities. Proteases are usually applied in much smaller quantities than the protein to be digested (10-1000 : 1) 34,35 to minimize self-digestion. We therefore performed a digestion on HRP pretreated with RapiGest™, DTT and IAM using only 10% proteinase K compared to HRP's mass.…”

Section: Resultsmentioning

confidence: 99%

Enzymatic digestion as a tool for removing proteinaceous templates from molecularly imprinted polymers

et al. 2017

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Section: Resultsmentioning

confidence: 99%

Enzymatic digestion as a tool for removing proteinaceous templates from molecularly imprinted polymers

et al. 2017

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“…One of the major limitations of PCR-based tests is inhibition of the nucleic acid amplification process by substances present in samples 35 . In urine, substances such as urea can act as PCR inhibitors, which can lead to inhibition of DNA polymerase or reverse transcriptase activity 36 , 37 .…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of BK virus detected in renal transplant patients in Sri Lanka: a preliminary study

Ratnayake

Fernando

Gajanayake

et al. 2022

Indian Journal of Medical Research

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Background & objectives: BK virus (BKV) is a polyomavirus and cause of a common infection after renal transplantation which could be preceded to BKV-associated nephropathy. It has four main subtypes (I–IV). BKV subtypes II and III are rare, whereas subtype I shows a ubiquitous distribution. The objective of the present study was to investigate the prevailing BKV subtypes and subgroups in renal transplant patients in Sri Lanka. Methods: The presence of BKV in urine was tested through virus load quantification by real-time PCR from 227 renal transplant patients who were suspected to have BKV infection. Of these patients only 41 were found to be BKV infected (>10 3 copies/ml) and those were subjected to conventional PCR amplification of VP1 gene followed by BKV genotyping via phylogenetic analysis based on DNA sequencing data. Results: Persistent BK viral loads varied from 1×10 3 to 3×10 8 copies/ml. Of the 41 patient samples, 25 gave positive results for PCR amplification of subtyping region of VP1 gene of BKV. BKV genotyping resulted in detecting subtype I in 18 (72%) and subtype II in seven (28%) patients. BKV subgroups of Ia, Ib-1 and Ib-11, and Ic were identified with frequencies of 6/18 (33.3%), 6/18 (33.3%), 5/18 (27.8%), and 1/18 (5.6%), respectively. Interpretation & conclusions: Findings from this preliminary study showed a high occurrence of subtype I, while the presence of subtype II, which is rare and less prevalent, was a novel finding for this Asian region. This emphasizes the need for further molecular and serological studies to determine the prevalence of different BKV subtypes in Sri Lanka.

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“…From our experience with various commercially available stool DNA extraction kits, the DNA extraction kit utilized in this study has good reliability and sensitivity. Previous studies have shown that treatment with proteinase K and heat may remove inhibitors from samples more efficiently than other treatment methods treatment (Bergallo et al, 2006;Schrader et al, 2012), which are part of the DNA extraction steps used in this study. Nonetheless, the use of internal/external controls and spike tests could be performed to confirm the absence of PCR inhibitors in the fecal samples in future studies.…”

Section: Discussionmentioning

confidence: 99%

A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces: development and validation

N.A.¹

2022

Trop Biomed

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Soil-transmitted helminth (STH) infections, mainly caused by Ascaris lumbricoides, Trichuris trichiura, and hookworms, are among the most common intestinal parasites that infect humans. The infections are widely distributed throughout tropical and subtropical countries, including Malaysia, particularly in underprivileged communities. Microscopic and culture techniques have been used as a gold standard for diagnostic techniques. However, these methods yield low sensitivity and specificity, laborious and time-consuming. Therefore, simple, rapid, and accurate alternative methods are needed for the simultaneous detection of STH infections. Although advanced technologies such as real-time multiplex PCR have been established, the use of this technique as a routine diagnostic is limited due to the high cost of the instrument. Therefore, a single-round multiplex conventional PCR assay for rapid detection of four STH species in the fecal sample was developed in this study. To perform the single-round multiplex PCR, each pair of species-specific primers was selected from target genes, including Ancylostoma duodenale (Internal Transcribed Spacer 2; accession No. AJ001594; 156 base pair), Necator americanus (ITS 2; accession No. AJ001599; 225 base pair), Ascaris lumbricoides (Internal Transcribed Spacer 1; accession No. AJ000895; 334 base pair) and Trichuris triciura (partial ITS 1, 5.8s rRNA and partial ITS 2; accession No. AM992981; 518 base pair). The results showed that the newly designed primers could detect the DNA of STH at low concentrations (0.001 ng/μl) with no cross-amplification with other species. This assay enables the differentiation of single infections as well as mixed infections. It could be used as an alternative and is a convenient method for the detection of STHs, especially for the differentiation of N. americanus and A. duodenale.

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Valutazione Della Carica Virale Del Citomegalovirus Umano (Hcmv)

Cited by 6 publications

References 0 publications

Enzymatic digestion as a tool for removing proteinaceous templates from molecularly imprinted polymers

Enzymatic digestion as a tool for removing proteinaceous templates from molecularly imprinted polymers

Molecular characterization of BK virus detected in renal transplant patients in Sri Lanka: a preliminary study

A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces: development and validation

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