Mucormycosis is difficult to diagnose. Samples from suspected cases often fail to grow Mucorales in microbiologic cultures. We identified all hematologic malignancy and stem cell transplant patients diagnosed with proven mucormycosis between 2001 and 2009 at Brigham and Women's Hospital/Dana-Farber Cancer Institute. Seminested PCR targeting Mucorales 18S ribosomal DNA and sequencing were perfor...

OBJECTIVE To identify adenovirus in patients with conjunctivitis by molecular methods such as the Polymerase Chain Reaction (PCR) and DNA sequencing. METHODS Samples of scrapings from the inferior fornix of 51 patients (39 diagnosed with Follicular Conjunctivitis and 12 diagnosed with Vernal Conjunctivitis) were processed by generic PCR to identify adenovirus. All the samples that were PCR po...

Background: Osteoporosis is a multifactorial disease. Mutation in cystein-rich domain2 (CRD2) and domain3 (CRD3) of osteoprotegrin protein can prevent its interaction with receptor activator nuclear factor kappa beta (RANKL) ligand lead to osteoporosis. Objectives: This study aimed investigate the possible alteration exon2 gene encoding CRD2 CRD3 domains osteoporotic women Chahar Mahal va Bakht...

One of the limitations of PCR to analyse a region of interest is that it requires knowledge of the flanking sequences in order to design amplification primers. Several adaptations of PCR have been described which allow some unknown sequences adjacent to regions of known sequence to be amplified. Primers directed to repetitive sequences, for instance Alu repeats can be used to pair with a primer...

The introduction of molecular detection technique namely the Sanger sequencing in the year 1977 made sequencing the order of nucleotides in a DNA molecule possible. It made a vast difference in the field of biology as the sequence abnormality detection in genetic disorders became much easier. In the year 1983, PCR technique was developed. And, application of it in a wide field of biology was ea...

Direct sequencing of PCR products by T7 DNA polymerase is prone to problems in GC-rich regions, due to the stability of secondary structure and rapid reannealing of the heat-separated complementary strands of DNA. To overcome these difficulties, we examined the effect of adding formamide to the sequencing reaction. Formamide weakens the hydrogen bonding between nucleotides (1, 2), and is, there...

Bisulfite sequencing is a key methodology in epigenetics. However, the standard workflow of bisulfite sequencing involves heat and strongly basic conditions to convert the intermediary product 5,6-dihydrouridine-6-sulfonate (dhU6S) (generated by reaction of bisulfite with deoxycytidine (dC)) to uracil (dU). These harsh conditions generally lead to sample loss and DNA damage while milder conditi...

Dactylogyrus spp. are monogenean worms found mostly as ectoparasites on the gills of several fish species, including carp and goldfish. These parasites are commonly detected by microscopic analysis of the gill lamellae, but this is time-consuming and technically difficult. In contrast to this conventional method, molecular techniques provide specific, sensitive and safe detection of parasites. ...

background: dna vaccination with plasmid encoding bacterial, viral, and parasitic immunogens has been shown to be an attractive method to induce efficient immune responses. bacteria of the genus brucella are facultative intracellular pathogens for which new and efficient vaccines are needed. methods: to evaluate the use of a dna immunization strategy for protection against brucellosis, a plasmi...

Damage to the genome is implicated in the progression of cancer and stress-induced diseases. DNA lesions exist in low levels, and cannot be amplified by standard PCR because they are frequently strong blocks to polymerases. Here, we describe a method for PCR amplification of lesion-containing DNA in which the site and identity could be marked, copied and sequenced. Critical for this method is i...