We develop a strategy for haplotype analysis of PCR products that contained two adjacent heterozygous loci using sequencing with specific primers, allele-specific primers, and ddNTP-blocked primers. To validate its feasibility, two sets of PCR products, including two adjacent heterozygous SNPs, UGT1A1⁎6 (rs4148323) and UGT1A1⁎28 (rs8175347), and two adjacent heterozygous SNPs, K1637K (rs1117601...

BACKGROUND Multilocus PCR coupled with electrospray ionization mass spectrometry (PCR/ESI-MS) is a new strategy for pathogen identification, but information about its application in fungal identification remains sparse. METHODS One-hundred and twelve strains and isolates of clinically important fungi and Prototheca species were subjected to both rRNA gene sequencing and PCR/ESI-MS. Three regi...

background: emergence klebsiella pneumoniae resistant to quinolone antibiotics due to mutations in gyra and parc genes created problem for treatment of patients in different hospitals in iran. the objective of this study was to determine the amino acid substitutions of gyra and parc proteins in certain clonal lineages of the k. pneumoniae conferring high level quinolone resistance. methods: one...

Amplicon sequencing utilizing next-generation platforms has significantly transformed how research is conducted, specifically microbial ecology. However, primer and sequencing platform biases can confound or change the way scientists interpret these data. The Pacific Biosciences RSII instrument may also preferentially load smaller fragments, which may also be a function of PCR product exhaustio...

Somatic mutation in human epidermal growth factor receptor-related 2 gene (HER2) is one of the driver mutations in lung cancer. HER2 mutations are found in about 2% of lung adenocarcinomas (ADCs). Previous reports have been based mainly on diagnostic screening by Sanger sequencing or next-generation sequencing (NGS); however, these methods are time-consuming and complicated. We developed a rapi...

An efficient approach for structural studies on bacterial chromosomes is presented. It is based on high-resolution PCR map construction by using a multiplex long accurate PCR (MLA PCR) protocol and a YAC clone carrying the region to be mapped as indicator. The high-resolution PCR map of the bacillus subtilis rrnB-dnaB region is presented as an example. Data are also presented on the use of DNA ...

the identification of mycobacteria in the species level has great medical importance. biochemical tests are laborious and time-consuming, so new techniques could be used to identify the species. this research aimed to the comparison of biochemical and sequencing 16s rdna gene methods to identify nontuberculous mycobacteria in patients suspected to tuberculosis in golestan province which is the ...

Background: Currently, it appears that new molecular-based methods could substitute microscopic and culture assessment for the first-line detection of microorganisms isolated from clinical specimens. However, will remain "continual strategy" until this technology is attuned to identifying all fungi can be biological Objectives: The present study aimed validate a high-resolution melting (HRM) te...