ABSTRACT
Calcifying nanoparticles are different forms of calcium and phosphate in sediments. Recent evidence suggests that calcifying nanoparticles [CNPs] are probably self-replicating. Several diseases are linked to nano-bacteria including kidney stones, gallbladder stone, cardiovascular plaques, oral-dental plaque, many cancers, and autoimmune diseases. The aim of this study was to assess the presence of nano-bacteria in kidney stones, gallbladder stones, and atherosclerosis plaques and compare them with each other in terms of structural form. In this study, 18 samples of kidney stones, 15 gallbladder stones, and 11 samples of atherosclerosis plaque, taken by surgeons during surgery, were collected. After isolation of nano-bacteria [NB] from the sample and confirmation by scanning electron microscope [SEM], some of the samples were cultured in a solution of Dulbecco's Modified Eagle Medium [DMEM] containing fetal bovine serum [FBS] 10% and glutamate at 37 °C with 5% CO[2] and 95% air for 8 weeks. Deoxyribonucleic acid [DNA] extraction protocol was performed in all samples. After that, they were analyzed by polymerase chain reaction [PCR] techniques. 17 cases out of 18 kidney stone samples were culture positive. Moreover, 14 cases out of 15 gallbladder stone samples and all 11 samples of atherosclerosis plaque were culture positive. Growth and the presence of CNPs were confirmed in all cases by SEM. PCR testing was performed for all samples; however, no DNA samples were detected with the primers used. According to the results, the presence of calcifying nanoparticles using culture methods, and observation using scanning electron microscope were confirmed. However, the existence of nucleic acid has not been confirmed in this study. Thus, more research using PCR must be performed to prove their existence. Therefore, we can say that one of the important factors in the development of kidney stones, gallbladder stones, and atherosclerosis plaques is the presence of microorganisms
ABSTRACT
Type 1 and type 2 herpes simplex [HSV] virus cause infection of central nervous system [encephalitis] in human. The molecular techniques are the best methods for detection of HSV. In this study we evaluated the novel molecular technique of LAMP for detection of HSV-1 and HSV-2. In this experimental study 184 cerebrospinal fluid [CSF] samples were collected from Mofid Hospital. DNA of every sample was extracted by use of Sinagen DNP kit. Based on the HSV DNA polymerase gene, a set of 6 primers were designed and sensitivity and specificity of this method were determined. By adding SYBER Green, LAMP product was identified. The results of LAMP method were compared to those of PCR by chi-square test. Sensitivity of LAMP method determined to be 5 copies/ tube and sensitivity of PCR method determined to be 50 copies/ tube. Both LAMP and PCR methods showed 100% specificity for detection of HSV type 1 and type 2. Among 184 samples, 60 samples were positive by LAMP but 45 samples were positive by PCR method. Sensitivity of LAMP was 10 times higher than that of PCR. Comparison of the results of the two methods by means of chi-square test showed a significant difference [p<0.05]. LAMP method had high sensitivity and specificity for detection of type 1 and type 2 HSV in CSF samples