Performance of the Enzyme Linked Fluorescent Assay in Diagnosis of HIV Infection; Comparative Evaluation with other Methods.

BACKGROUND: Our study aimed to investigate the role of the enzyme linked fluorescent assay (ELFA) method in the diagnosis of Human Immunodeficiency Virus (HIV) infection by comparing it with enzyme-linked immunosorbent assay (ELISA) and chemiluminescent microplate immunoassay (CMIA) methods and its role in the HIV diagnostic algorithm and to update the recommended algorithm for HIV testing. METHODS: We evaluated 101 HIV-reactive and 101 HIV-negative specimens. All samples were studied with the methods of anti HIV1/2 test micro-ELISA, ELFA, and CMIA. At the same time, HIV RNA PCR and western blot (WB)/rapid immunochromatographic test (RICT) were also studied with the same samples. RESULTS: All HIV RNA and WB positive samples (n = 101) were positive with micro-ELISA, CMIA and ELFA. Twenty-five negative samples of HIV RNA and WB were positive with micro-ELISA and CMIA, while just 6 samples were positive with ELFA. When all samples were evaluated together, the false positivity rate of the ELFA method was found to be 5.9%, and the false positivity rates of the micro-ELISA and CMIA methods were determined to be 31.7% and 30.7%, respectively. CONCLUSIONS: It was determined that there is a high level of agreement between the ELFA method and confirmation tests. It was thought that it might take place in the preconfirmation stage. As can be seen from the results obtained, the false positive rate by ELFA method was found to be about five times lower than that of micro-ELISA and CMIA methods. Considering that antigen (p24) and antibody positivity can be given separately with this aspect, it can be considered that there is a confirmation place in HIV diagnosis algorithm.

Distribution of respiratory viruses: Evaluation of multiplex PCR results of 3074 patients.

OBJECTIVE: Early and accurate diagnosis of acute respiratory infections is important because these diseases negatively affect public health and can lead to loss of workforce and an increase in health expenditures. In this study, we aimed to determine the respiration panel multiplex polymerase chain reaction (PCR) test results and seasonal distribution in our region. METHODS: Three thousand and seventy-four patients samples multiplex PCR (Anatolia, Bosphore® Respiratory Pathogens Panel Kit v1) test results, which were sent to our laboratory, from 13 hospitals in our region between January 2018 and December 2018, were evaluated retrospectively. RESULTS: A total of 3074 patients samples, 1465 (48%) were positive and 1609 (52%) were negative test results. The most common factors were rhinovirus 30.2%, influenza A 23.1%, and respiratory syncytial virus (RSV) A/B 19.1%, respectively. When the distribution of these three most common viruses by months is examined, the most frequent months were determined as June for rhinovirus, November for influenza A, and February for RSV A/B. In the period between October and February, there was a significant increase in the positivity level of viral factors. CONCLUSION: The use of molecular methods in the diagnosis of respiratory infections will prevent unnecessary use of antibiotics and ensure correct and rapid treatment.

A Predictive Test in The Screening of HIV Positive Cases; Interferon Gamma Inducible Protein 10 (IP-10).

BACKGROUND: HIV (human immunodeficiency virus), causing acquired immunodeficiency syndrome (AIDS), is one of the most important health problems in the world. Certain cytokines produced during the cytokine storm in an acute infection can be biomarker candidates. The strong association of IFN-γ inducible protein 10 (IP-10) with low CD4 cell counts suggests that it can be an acute phase biomarker. METHODS: In this study, IP-10 was monitored at routine controls during pre-treatment and/or in subsequent phases of treatment, and its correlation with CD4 cell count and viral load was assessed. Venous blood samples, taken from 30 patients (at 0 - 3 - 6 months), and 20 healthy volunteers, were sent to the Laboratory for flow cytometry, nucleic acid tests (NAT) and ELISA tests. RESULTS: The mean IP-10 concentration of patients was 344 pg/mL, and these values for the untreated, treated and control groups were 422 pg/mL and 210 pg/mL and 68 pg/mL respectively. A statistically significant difference was found between the IP-10 values of the patient and control groups (p = 0.006). There was a significant, positive and moderate relation between IP-10 and viral load values (r = 0.59, p < 0.001), while there was a significant, negative and moderate relation between IP-10 and CD4 cell count (r = -0.51, p < 0.001). CONCLUSIONS: IP-10 levels in early HIV-1 patients, which are shown to be closely related to CD4 cell levels and viral replication, may be an alternative or support marker compared to the more expensive viral load tests in monitor-ing viremia changes and response to antiretroviral treatment.

The role of G protein-associated estrogen receptor (GPER) 1, corin, raftlin, and estrogen in etiopathogenesis of intrauterine growth retardation.

Objective: The aim of the present study was to detect the role of G protein-associated estrogen receptor (GPER) 1, corin, raftlin and estrogen in etiopathogenesis of intrauterine growth retardation (IUGR).Materials and methods: The present study was designed prospectively between January 2017 and May 2018. The study group included 32 patients with unexplained IUGR and 32 healthy pregnant women who gave birth at term among the patients who referred to obstetrics clinic of a tertiary reference hospital. Intrauterine growth retardation (IUGR) was accepted as birth weight below 10th percentile according to the estimated fetal weight. Exclusion criteria were as follows: the patients with renal or hepatic dysfunction, presence of any chronic disease, smoker patients, preeclampsia, acute or chronic inflammatory diseases, body mass index as <18 kg/m2 and >25 kg/m2, structural or chromosomal abnormality in fetus Estradiol (E2), estriol (E3), GPER, corin, and raftlin levels were analyzed in maternal serum and placental tissue homogenate through ELISA method.Results: Serum levels of GPER-1, raftlin, and E3 were significantly lower in IUGR group when compared with the control group (p < .05 for all). Serum corin and E2 levels were similar between two groups. GPER-1, E2, E3, raftlin, and corin levels in placental homogenate were found significantly higher in the control group (p < .05 for all).Conclusion: Although maternal, fetal, and placental causes take place in etiopathogenesis of IUGR, exact etiological factor is not revealed in majority of the IUGR cases. The present study serves as the first study revealing the role of the decrease in GPER-1 and raftlin in maternal serum and placental levels on the etiopathogenesis of IUGR. Furthermore, the decrease in placental corin expression of the cases with IUGR was detected first in the literature. The present study reveals a potential therapeutic use of GPER-1, corin, and raftlin for IUGR.

[Employee Safety and Biosafety Applications in Microbiology Laboratories; A Multicenter Evaluation in Turkey]. / Mikrobiyoloji Laboratuvarlarinda Çalisan Güvenligi ve Biyogüvenlik Uygulamalari Açisindan Türkiye'de Çok Merkezli Bir Degerlendirme.

Medical laboratory personnel may be exposed to various hazards, especially biological and chemical, during their routine activities. In this multicenter study, which could reflect the nation wide results, it was aimed to determine the safety and biosecurity practices of the employee working in medical microbiology laboratories and to reveal the current situation. A total of 1072 personnel working in the Medical Microbiology Laboratory of 23 hospitals (14 medical faculty hospitals, seven ministry of health training and research hospitals and two state hospitals) from different provinces were provided with a questionnaire consisting of 33 questions inquiring about the rules, opinions, attitudes and behaviors regarding safety and biosafety practices. Statistical analyses were made with institutions, age groups, gender, educational background, working time and occupational groups in terms of exposure to biological and chemical hazards. It was determined that approximately 50% personnel of the university/ training and research hospitals and 2/3 of the state hospitals personnel consumed food and beverages in the laboratories (p<0.05). Compared with other hospitals, it was determined that in state hospitals; the absence of separate resting room (35%), the personnel finding their own knowledge and practices inadequate (28.9%), laboratory coats washed at home (95%), educational organization and participation rates (90%) and medical waste information levels of the personnel were higher (p< 0.05). It was determined that as the age progresses, the rate of education, food and beverage consumption in the laboratory, not being outside the laboratory with protective equipment (gloves, masks and laboratory coats) and the history of laboratory acquired infections were increased (p< 0.05). It was observed that washing the laboratory coats at home was higher in the younger age group and hospital washing was higher in the elderly group (p< 0.05). There was no significant difference between the genders in terms of food and beverage consumption in the laboratory (p= 0.09). It was determined that periodic health checks were not performed in 1/3 of both sexes, but the use of gloves and compliance with medical waste rules was lower in men. Female employees find themselves inefficient in terms of knowledge and practices (p< 0.05). The rate of those who did not have their periodic checkups at regular intervals was higher in the high school and master of science education groups; While non-compliance with medical waste rules, food and beverage consumption in the laboratory was highest in the primary and high school graduates, the lowest rates were found in the master and doctorate groups (p< 0.05). The rate of those who had regular health checkups was higher in the group of specialist physicians and technicians (p< 0.05). It was observed that the rule of not going out of the laboratory with protective equipment was fully observed in the 35+ years working group, while compliance was 70-85% in other groups (p< 0.05), hepatitis B vaccination rate was highest in specialist doctors and lowest in cleaning and other personnel group (p< 0.05). Highest non-compliance rate with medical waste rules was observed in the cleaning personnel group (p< 0.05). As a result, although advances have been made in employee safety practices in medical microbiology laboratories in our country in recent years, it has been found that it is not yet sufficient. The results indirectly reflected the profile of medical laboratories in our country. In the laboratories, physical space and equipment deficiencies should be eliminated, periodic health checkups and vaccination should be provided, non-staff entrance to the laboratory and food, beverage and cigarette consumption should be prevented, laboratory coats should be washed in the hospital, in-service trainings, including medical waste training, should be conducted and these trainings should be developed through mechanisms that will change the behavior.

[Comparison of Antiviral Effect of Olive Leaf Extract and Propolis with Acyclovir on Herpes Simplex Virus Type 1]. / Zeytin Yapragi Ekstresi ve Propolisin Herpes Simpleks Virüsü Tip 1 Üzerine Antiviral Etkisinin Asiklovir ile Karsilastirilmasi.

While acyclovir, a nucleoside analogue, is widely used for herpes simplex virus type 1 (HSV-1), emergence of drug-resistant viruses due to frequent usage of this class of medicines, and their toxic side effects require exploring novel active molecules. Despite the studies on developing synthetic molecules in medical sciences and pharmacology, herbs as a natural source of biologically-active compounds remain popular. In this in vitro study, olive leaf extract (OLE) and propolis alone or in combination with acyclovir were investigated for their antiviral efficacy in HSV-1.Toxic doses of OLE, propolis, and dimethyl sulfoxide, propolis diluent, for Hep-2 (ATCC, CCL-23) cells were determined by conventional cell culture. Using "endpoint" method, the viral dose infecting half of the cell culture (TCID50) was calculated, and viral quantity was determined with Spearman-Karber method. Antiviral effects of OLE and propolis on HSV-1 were investigated by conventional cell culture and real-time cell analysis (RTCA). Combinations of the two extracts with one another and with acyclovir were evaluated by RTCA. Active substances prepared at three different dilutions were added to tubes with HSV-1 of logTCID50: 11.5 in descending order starting from the highest non-toxic concentration, and they were left at room temperature for two different durations (one hour and three hours). The aliquots taken from the tubes were cultured in plates containing Hep-2 cells and evaluated after 72 hours. Combinations of extracts and acyclovir at concentrations at least four times lower than the lowest concentration showing antiviral efficacy against HSV-1 were cultured with Hep-2 cells in the e-plates of the xCELLigence RTCA device, measurements were obtained at 30 minute intervals, and data were recorded in real time. In the test with two different durations and at different concentrations of OLE and propolis, antiviral efficacy was observed both with one-hour and three-hour incubation at a concentration of 10 µg/ ml for propolis and 1.2 mg/ml for OLE with RTCA. The duration and concentration of the greatest decrease in viral quantity were in the first one hour and 10 µg/ml for propolis, and in the first one hour and 1.2 mg/ ml for OLE. Combination of propolis and OLE with acyclovir caused no cytopathic effects, and the combination of extracts led to delayed cytopathic effect. According to these results, propolis and OLE, alone and in combinations with acyclovir, have antiviral efficacy against HSV-1. These agents may reduce the dose and side effects of acyclovir in case of co-administration since they exert their effects through a different mechanism than acyclovir,possibly through direct virucidal activity, inhibition of virus internalization or viral inhibition in early stages of replication (inhibition of adsorption/binding of viral particles to the cell). These extracts that do not require conversion to active form have the potential to reduce infectivity in oral lesions, prevent spread, and be used in the topical treatment of acyclovir-resistant HSV infections, particularly in immunocompromised patients. However, in vivo studies should be conducted to determine their medicinal properties and potential toxicities. These results should be supported by further comprehensive studies and the efficacy against other viruses should also be investigated.

[Evaluation of the approaches and knowledge level of tuberculosis diagnosis and treatment of infectious disease research assistants and specialist physicians]. / Infeksiyon hastaliklari ve klinik mikrobiyoloji arastirma görevlisi ve uzman hekimlerinin tüberküloz tani ve tedavisinde yaklasimlarinin ve bilgi düzeylerinin degerlendirilmesi.

INTRODUCTION: This study aims to evaluate approaches and knowledge level for tuberculosis (TB) diagnosis and treatment among infectious diseases and clinical microbiology research assistants and specialist physicians. MATERIALS AND METHODS: This was a descriptive study on the research assistants and specialist physicians. A questionnaire consisting of 24 questions prepared by the researchers was used. Data were analyzed using SPSS, version 22.0. RESULT: In this study, 116 physicians participated. The average age of participants was 41.07 ± 8.65 years, and 64.7% were female. The proportion of physicians with no pulmonary and non-pulmonary TB experience was calculated as 6.9% and 3.4%, respectively. Acid-fast-bacilli (AFB) tests were most frequently used in the diagnosis of pulmonary TB. On the other hand, the tuberculin skin test (TST) + chest radiography (CR) was most frequently used for latent TB in immunosuppressed subjects (91.4% and 69%, respectively). The most common non-pulmonary TB clinic form encountered was lymphadenitis (88.8%); the most common treatment problem was drug side effects (75%); and the most common drug resistance was to isoniazid (25.9%). It was determined that physicians encountered HIV and TB coinfection in 62.9% of patients, and standard TB treatment was applied for most patients (48.3%). The most frequent yearly TST + CR + AFB trials (69%) were performed in the screenings of health workers who were in contact with TB patients. It was observed that 44% of physicians used a negative pressurized chamber and/or isolation chamber in the institution, and 72.4% used a N95/FFP3 mask during examination. It was determined that the participants were mostly correct about TB. CONCLUSIONS: Infectious diseases and clinical microbiology physicians continue to encounter TB patients. This group of physicians often has a good level of knowledge and experience with TB, although special training for this disease is low.

Increased oxidative stress is associated with insulin resistance and infertility in polycystic ovary syndrome.

OBJECTIVES: The present study aims to investigate the role of oxidant-antioxidant status in young women with polycystic ovary syndrome (PCOS). MATERIAL AND METHODS: Seventy-one women with PCOS and 53 healthy controls are compared in aspect of demographic characteristics, biochemical data, hormones, and oxidant-antioxidant status. RESULTS: The PCOS group had significantly lower zinc, higher malondialdehyde and gluthathione peroxidase and lower serum catalase levels than the control group (p = 0.016, p < 0.001, p = 0.043 and p = 0.025 respectively). The PCOS patients with IR had significantly higher malondialdehyde, lower catalase and serum zinc levels than the PCOS patients without IR (p = 0.015, p = 0.010, p = 0.001 respectively). The infertile PCOS patients had significantly higher malondialdehyde, lower catalase and serum zinc levels than the fertile PCOS patients (p = 0.022, p = 0.045,p = 0.001 respectively). There was a statistically significant and positive correlation between HOMA-IR and malondialdehyde values (r = 0.523, p = 0.001), between HOMA-IR and glutathione peroxidase values (r = 0.468, p = 0.001) and between HOMA-IR and zinc values (r = 0.601, p = 0.001). There was a statistically significant and negative correlation between HOMA-IR and catalase values (r = -0.493, p = 0.001). CONCLUSIONS: The patients with PCOS are under oxidative stress and this oxidative stress seems to be the highest in patients with IR and with infertility. Despite the prominent increase in the oxidative stress, there was a variation in the antioxidant response.

[Investigation of human papillomavirus prevalence in women in Eskisehir, Turkey by Pap smear, hybrid capture 2 test and consensus real-time polymerase chain reaction and typing with pyrosequencing method]. / Eskisehir'de kadinlarda insan papillomavirus prevalansinin, Pap yaymasi, hibrid yakalama 2 testi ve konsensus gerçek zamanli polimeraz zincir reaksiyonu ile arastirilmasi ve pirodizileme yöntemiyle tiplendirilmesigi.

Human papillomavirus (HPV) infections have a broad range of clinical spectrum from subclinical or asymptomatic infection to anogenital carcinoma. The detection of HPV-DNA and determination of the risk groups in cervical cancer (CC) screening is very important because CC is considered to be a preventable illness which is the third most common cancer type of women in the world. The aims of this study were to investigate the presence of HPV-DNA in women by two different molecular methods and to compare their results together with the results of cytology, in Eskisehir, Central Anatolia, Turkey. A total of 1081 women aged between 30-65 years, who applied to Eskisehir Early Diagnosis, Screening and Training of Cancer Center (KETEM) for screening were included in the study. Three separate cervical samples were collected simultaneously from the participants for cytologic examination and molecular studies. In the first step of the study, all cervical samples were investigated for the presence of HPV-DNA by Hybrid Capture 2 (HC2; Qiagen, Germany) method. In the second part of the study, consensus real-time polymerase chain reaction (RT-PCR) (Takara Bio Inc., Japan) was performed in 152 samples which included HC2 positive and randomly selected negative samples, and then the HPV genotypes were detected by using a commercial kit based on pyrosequencing method (Diatech Pharmacogenetics S.R.L, Italy). In the first part of the study, HC2 test was found positive in 3% (32/1081) of the women, while in 4.4% (47/1081) Pap smear was positive alone or with HC2 test. Five (0.5%) samples yielded positive results with both of the methods, and four of them were positive for high risk HPV types. Cytology results were negative in 19 out of 23 (23/1081, 2.1%) samples that were reported as high risk HPV by HC2 test. On the other hand, 42 (42/1081, 3.9%) samples that were positive by cytology yielded negative results by HC2 test. In the second part of the study, 32 (21.1%) of 152 selected samples were positive by HC2 test, 40 (26.3%) were positive by Pap smear, and 53 (34.9%) were positive by consensus RT-PCR. All of the 32 samples that were positive by HC2 were also positive by RT-PCR, however 21 samples that were positive by RT-PCR were negative by HC2 test. Among 40 samples that were positive (abnormal) by Pap smear, HPV-DNA was positive in nine (22.5%) by RT-PCR and in five (12.5%) by HC2 test, but HPV-DNA was not detected in 31 (77.5%) samples by both of the tests. Genotyping of the strains could be performed in 44 samples, and the most common type detected was HPV type 16 (n=15, 34.1%), followed by type 90 (n=11, 25%) and type 18 (n= 4, 9.1%). In our study, the sensitivity, specificity, positive and negative predictive values of Pap smear method were estimated as 16.1%, 96%, 10.6% and 97.5%, respectively, based on the HC2 results which was approved by U.S. Food and Drug Administration (FDA). In addition, a significant degree of concordance was detected between HC2 and concensus RT-PCR methods (Cohen's kappa: 0.665). In conclusion, regarding the insufficient number of cytopathologists in our country and according to the recommendations of American Society for Colposcopy and Cervical Pathology (ASCCP) and FDA, it was once again demonstrated that, the implementation of molecular diagnostic methods in addition to the Pap smear for effective screening of CC are needed.