Polymerase Chain Reaction Sequencing Techniques Research Articles

Assessment of NLRP3 Gene Polymorphisms with Periodontitis as Compared with Healthy Periodontium in Iraqi Arabs Patients.

The nod-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome regulates the maturation and release of the cytokines as well as the activation of caspase in response to danger signals derived from pathogenic infection, tissue damage, andmetabolic changes that have a role in the pathogenesis of different diseases as periodontitis. Yet, the susceptibility to this illness could be determined by population-based genetic differences. The aim of this study was to determine whether periodontitis in Arab populations from Iraq is correlated with NLRP3 gene polymorphisms and measure clinical periodontal parameters and investigate their association with genetic polymorphisms of the NLRP3. The study sample consisted of 94 participants ranging from 30 to 55 years old, both males and females who fulfilled the study's criteria. The selected participants were divided into two groups: the periodontitis group (62 subjects) and the healthy control group (32 subjects). The examination of clinical periodontal parameters of all participants was carried out, followed by a collection of venous blood for NLRP3 genetic analysis using the polymerase chain reaction-sequencing technique. The genetic analysis of NLRP3 genotypes at four single nucleotide polymorphisms (SNPs) (rs10925024, rs4612666, rs34777555, and rs10754557), by Hardy-Weinberg equilibrium, identified nonsignificant differences in studied groups. The C-T genotype among periodontitis was significantly different from controls, while the C-C genotype among control was significantly different from periodontitis at NLRP3 rs10925024. Overall, there were 35 SNPs in the periodontitis group and 10 SNPs in the control group for rs10925024 with significant differences versus nonsignificant differences of the other SNPs between the studied groups. Clinical attachment loss and NLRP3 rs10925024 additionally demonstrated a significant positive correlation in the periodontitis subjects. The findings suggested that polymorphisms of the NLRP3 gene may have a role and increasing the genetic susceptibility to periodontal disease in Arabs Iraqi patients.

Microsporidia and Coccidia as Causes of Persistence Diarrhea Among Liver Transplant Children

We determined species/genotype(s) of enteric microsporidia and coccidia causing diarrhea among 44 liver transplant children in Shiraz Nemazee hospital using acid-fast-trichrome staining and polymerase chain reaction-sequencing techniques. Enterocytozoon bieneusi (genotype D), Cryptosporidium (parvum and meleagridis) were detected in 6.81% and 11.36% of the children, respectively.

Detection of Mycobacterium kansasii infection in a rhesus macaque ( Macaca mulatta) using a modified QuantiFERON-TB Gold assay

A modification of a highly practical human interferon-gamma release assay for the diagnosis of tuberculosis (TB), the QuantiFERON ®-TB Gold (In-Tube Method) (QFG-IT) assay, was evaluated for diagnosing natural mycobacterial infection in rhesus macaques ( Macaca mulatta). All animals in a captive colony were tested using the QFG-IT and tuberculin skin test (TST). Animals testing positive to these tests were euthanised and necropsied. Selected tissues were processed for histopathology and mycobacterial culture, and positive cultures were speciated by Mycobacterium tuberculosis complex polymerase chain reaction (PCR) and 16S rRNA gene PCR sequencing techniques. M. tuberculosis was cultured from a TST-positive/QFG-IT-positive animal which showed gross pulmonary pathology typical of TB. Additionally, Mycobacterium kansasii was cultured from a TST-negative/QFG-IT-positive animal which had no pathological or histopathological signs of mycobacterial infection. The detection of M. kansasii infection in a QFG-IT-positive animal which showed no evidence of disease indicates that this test might be a highly sensitive tool for the diagnosis of mycobacterial infection in rhesus macaques. However, these findings highlight the limitations of the QFG-IT to specifically detect infection by the pathogens M. tuberculosis and Mycobacterium bovis.