The investigation of community-acquired and nosocomial respiratory syncytial virus and other viral respiratory tract infections in children.

Although respiratory viruses are known as the major causes of community-acquired respiratory tract infections all over the world, they can also cause serious nosocomial respiratory infections and hospital outbreaks. The aim of this study is to investigate the incidence of community-acquired and nosocomial RSV and other viral respiratory tract infections in children hospitalized at the Pediatric Intensive Care Unit of Cukurova University Faculty of Medicine. Nasopharyngeal swab samples were taken from 100 children aged 0-16 years with suspected community-acquired (60) and nosocomial (40) respiratory tract infections from September 2018 to June 2021. The Multiplex real-time PCR test was used for the diagnosis of respiratory viruses. Of children with community-acquired respiratory tract infections, 65% (39/60) were positive for at least one virus and the rate of coinfection in this group was 35.9% (14/39). In children with nosocomial respiratory tract infection, positivity was detected to be 62.5% (25/40) and the coinfection rate was 40% (10/25). The most predominant virus in community-acquired respiratory tract infections was influenza A virus (25%), followed by ADV (18.3%), hBoV (15%), RSV (11.7%), and RhV (10%). In nosocomial viral respiratory tract infections, the most common virus was RSV (20%), followed by influenza A virus (12.5%), RhV (12.5%), ADV (12.5%), hMpV (10%), and hBoV (10%). Early diagnosis of respiratory viral infections with real-time PCR test is important in terms of reducing morbidity and mortality, applying control methods to prevent the spread of nosocomial viruses, shortening the hospitalization period, preventing the use of unnecessary antibiotics, and giving appropriate antiviral treatment.

BK Virus Infections and Hemorrhagic Cystitis in Allogeneic Hematopoietic Stem Cell Transplant Recipients.

BK virus (BKV) associated with hemorrhagic cystitis (HC) is the most important complication that develops after hematopoietic stem cell transplantation (HSCT) in patients with hematological malignancies. This study aims to investigate BKV infections and HC in pediatric patients after allogeneic hematopoietic stem cell transplantation. Between November 2018 and November 2019, a total of 51 patients between the ages of 11 months and 17 years were included in the study. BKV Bosphore ® v1 quantification kit (Geneworks Anatolia, Turkey) was used for the detection of BKV DNA in urine and blood samples. Among the total of 51 patients, the incidence of BKV infection was found to be 86.3%. Allogeneic HSCT was performed in 40 patients and autologous HSCT in 11 patients. BK viruria and/or viremia were detected in 85% (44) of patients who underwent allogeneic HSCT and in 90% in the autologous group. High-level BK viruria (>107 copies/mL) was found in 41% (9) of 22 patients who were BKV positive before transplantation, while in 27.5% (8) of 29 patients who were BKV negative before transplantation; thus, BKV positivity before transplantation was considered a risk factor for high-level BK viruria. Acute GVHD developed in 6 of 40 patients in the allogeneic group. HC was prevented in 12 (67%) of 18 patients who received preemptive treatment, while HC developed in 6 (33%). HC occurred at a median of 35 days (17-49 days) post-transplant. Despite preemptive treatment, 6 (15%) patients who developed HC associated with BKV were in the allogeneic group but not in the autologous group. Of these patients with HC, 5 received a myeloablative treatment regimen, and 1 patient was given a reduced-intensity treatment regimen. The viral load in urine was found to be 107-9 copies/mL within 2 weeks before the development of HC and has been identified as a prognostic indicator. In conclusion, early diagnosis of viral infections by monitoring BKV viral load in HSCT patients will be effective in preventing the progression of complications such as BKV-associated HC by providing timely initiation of preemptive treatment.

In Vitro efficacy of cyclosporine a and various antiseptics and antiviral drugs on adenovirus genotype 8, a common cause of epidemic keratoconjunctivitis.

PURPOSE: To evaluate the in vitro efficacy of cidofovir, ganciclovir, povidone-iodine, chlorhexidine, and cyclosporine A on adenovirus genotype 8. METHODS: Conjunctival samples were collected from patients with adenoviral conjunctivitis and cultured in A549 cells. Adenovirus diagnosis was confirmed by RT-PCR. For each drug, the 50% cytotoxic concentration (CC 50 ) was determined. Subsequently, the antiviral activity was tested at concentrations below CC 50, and the 50% inhibitor concentration (IC 50 ) of drugs was determined RESULTS: While the IC 50 of cidofovir against adenovirus genotype 8 was 3.07 ± 0.8 µM, ganciclovir, povidone-iodine, chlorhexidine, and cyclosporine A were not found to be effective against adenovirus genotype 8 at concentrations below the CC 50 value. CONCLUSIONS: Cidofovir was found effective and the IC 50 value was within the ranges in the literature. Ganciclovir and cyclosporine A were found to be ineffective at doses below the cytotoxic dose, povidone-iodine and chlorhexidine was found to be highly cytotoxic.

The Relationship between Clinical Findings and Viral Load in Adenoviral Keratoconjunctivitis.

The aim of this study was to evaluate the relationship between clinical findings and viral load in adenoviral keratoconjunctivitis (Ad-Kc). In this cross-sectional study, 30 eyes of 30 patients with Ad-Kc were assessed. Real-time polymerase chain reaction was performed to detect and quantify adenovirus in all samples. Patients were divided into three subgroups according to baseline viral load (<107, 107-108, >108 human adenovirus [HAdV] copies/mL). The duration of follow-up, HAdV DNA copy number, treatment regimen, and detailed clinical findings, including uncorrected visual acuity, eyelid edema, conjunctival hyperemia, chemosis, follicular reaction, corneal involvement, conjunctival pseudomembrane, and subepithelial infiltrates (SEIs) were recorded. This study showed that a high initial viral load was associated with the development of SEIs and pseudomembrane formation (P < 0.05). The clinical findings and ocular complications of Ad-Kc were similar in the treatment groups at the final examination (P > 0.05). Our results show that a high initial viral load in Ad-Kc may be predictive of inflammatory sequelae. Determining the initial viral load in Ad-Kc may help understand the clinical course of the disease better and prevent complications.

Clinical aspects and prognosis of polymicrobial keratitis caused by different microbial combinations: a retrospective comparative case study.

PURPOSE: The purpose of this study is to compare the predisposing factors, clinical findings, treatment results, and prognosis for polymicrobial keratitis. METHODS: In this retrospective comparative case study, we identified the cases of polymicrobial keratitis from the microbiological records (n = 649) at Balcali Hospital, Çukurova University (Adana, Turkey; October 2010-2018). We included all the cases of infectious keratitis with two different types of microbial agents and grouped them as follows: group 1 (n = 25), bacterium-fungus coexistence; group 2 (n = 12), herpes simplex virus (HSV) or Acanthamoeba with bacterial infection; and group 3 (n = 7), HSV or Acanthamoeba with fungal infection. We compared the clinical and microbiological characteristics, and treatment outcomes among the groups. RESULTS: In our study, we found that 44 infectious keratitis cases (6.7%) were of polymicrobial nature. The mean follow-up period was 11.4 ± 17.8 months. In total, 17 different bacteria along with 3 different fungi, HSV, and Acanthamoeba were isolated. The most common bacterium was Staphylococcus epidermidis (25%). Most of the fungal pathogens were filamentous. Patients with initial treatment failure and requiring surgical intervention had larger infiltrates (p = 0.023, p = 0.003, respectively) than other patients. Older age was associated with delayed recovery and poor visual prognosis. CONCLUSIONS: Bacterial-fungus coexistence is the most common combination among patients, but other combinations should also be considered for suspected polymicrobial etiology. The corneal infiltrate size may be an important indicator of the course of disease and response to treatment. A closer and longer follow-up period should be planned for older patients.

Atypical herpes simplex keratitis: frequency, clinical presentations and treatment results.

AIMS: To determine herpes simplex virus (HSV) DNA positivity in corneal scraping samples obtained from patients with microbial keratitis whose findings were not specific for HSV keratitis and to evaluate these particular cases with respect to clinical features and antiviral treatment results. METHODS: Records of patients with microbial keratitis treated in a tertiary eye care hospital within the 3-year period were evaluated retrospectively. Real-time polymerase chain reaction (PCR) was used to identify HSV DNA. Smear slides were evaluated by light microscopy. Patients with typical presentations and histories of HSV keratitis were excluded. RESULTS: Two hundred and seventy-six eyes of 276 patients were included in the study. HSV-1 DNA was detected in 25 eyes (9%). In these 25 eyes, the initial diagnosis was fungal or bacterial keratitis. The mean symptom duration was 20 ± 14 days (2-60 days). The risk factors were ocular surgery (20%), blepharitis (16%), trauma (8%) and contact lens wear (4%); however, the majority of patients did not have any specific cause for keratitis (52%). Clinical features were variable and not typical for any particular etiology. Culture and microscopic examinations revealed bacteria and/or fungi in 6 patients in addition to herpes infection. Antiviral treatment was successful in 72% of patients. CONCLUSION: Herpetic corneal infections can present without typical dendritic or geographic ulcers and may be masked by other infections. Real-time PCR is a useful method for rapid and definitive diagnosis. HSV infection should be considered for microbial keratitis without specific risk factors, with negative culture results and poor response to antimicrobial agents.

[Detection of influenza virus infections by molecular and immunofluorescence methods]. / Influenza virüs enfeksiyonlarinin moleküler ve immün floresan yöntemlerle saptanmasi.

Influenza virus infections are extremely important for human health due to the occurence of seasonal epidemics and pandemics worldwide. Influenza is associated with high morbidity and may result in serious complications such as life threatening viral or bacterial pneumonia. Especially, young children, older adults, patients with chronic diseases such as heart, lung, kidney, and diabetes and immunosuppressed people are at higher risk for complications and death from influenza virus infections. The aim of this study was to determine the incidence of influenza type A and B virus infections and influenza A virus subtypes in hospitalized patients with respiratory tract infections by real-time reverse transcriptase-polymerase chain reaction (RT-PCR, Sacace, Italy), conventional RT-PCR and direct immunofluorescence antibody (DFA, Argene SA, France) tests. Nasopharyngeal swab specimens were collected from a total of 476 patients with respiratory tract symptoms by using flocked swabs (Copan Diagnostics, Italy) between 1 April 2012 and 31 December 2013. Influenza A virus was detected in 20.5% (98/476) and influenza B virus in 3.3% (16/476) of the cases by real-time RT-PCR test. During the study period, 63.3% of 98 influenza virus isolates were found as influenza A(H1N1)pdm09 and 36.7% were influenza A(H3N2) subtypes. Influenza A (H1N1) pdm09 subtype was observed in 12 cases in January 2013 and influenza A(H3N2) subtype was observed in 11 cases in December 2013 as the highest values. When the real-time RT-PCR test was regarded as the reference test, the sensitivities of DFA test for influenza A and B and conventional RT-PCR test with WHO primers (M30F2/08 and M264R3/08) for influenza A were detected as 72.4%, 75%, 96% and the specificities were detected as 99.2%, 99.5% and 100%, respectively. In conclusion, influenza A virus infection was detected rather high with a rate of 20.5% in the study group. The monitoring of influenza virus types and subtypes is required for the evaluation of influenza vaccine strains and circulating influenza viruses and for the identification of subtypes with pandemic potential. Planning for appropriate antiviral therapy using real-time RT-PCR in the early diagnosis of influenza virus infections will significantly contribute to the management of the patient's treatment. Thus, unnecessary drug use will be prevented and controlled with effective treatment of the disease at the time of infection.

Seroprevalence of parvovirus B19 in fibromyalgia syndrome.

This study was aimed to evaluate the seroprevalence of parvovirus B19 in patients with fibromyalgia syndrome (FS). Seventy-five patients with FS (44.3 +/- 8.3) and 75 healthy controls (44.2 +/- 8.1) were evaluated. Serum anti-B19 IgM and IgG antibodies were measured by ELISA technique. Patients were questioned about duration of symptoms, characteristic features of FS, and symptoms related with viral infection preceding the onset of FS. No significant difference was found regarding the prevalence of anti-B19 IgM antibodies between the groups (p = 0.494). Seropositivity of anti-B19 IgG of the patients was significantly higher than control group (81.3% vs. 64% respectively, p = 0.027). No statistically significant differences were found regarding to the clinical features between fibromyalgia patients with IgG antibody compared to those without IgG antibody. Parvovirus B19 IgG seropositivity was found to be significantly higher in patients with FS. Parvovirus B19 infection might have a role in the etiopathogenesis of FS or might act as a triggering factor.

Asymptomatic herpes simplex virus type 2 (HSV-2) infection among pregnant women in Turkey.

BACKGROUND & OBJECTIVES: A large proportion of individuals with serologic evidence of infection with herpes simplex virus type 2 (HSV-2) are asymptomatic. HSV-2 is the main cause of genital herpes infections. The acquisition of genital herpes during pregnancy has been associated with spontaneous abortion, premature labour and congenital and neonatal herpes. The present study was undertaken to determine asymtomatic genital HSV-2 shedding and seroprevalence of HSV-2 infection among asymptomatic pregnant women at the time of delivery in Adana, Turkey. METHODS: Asymptomatic 130 pregnant women without a history of genital herpes were enrolled in the study. HSV-2 shedding was determined by viral culture of the swabs collected from cervix and vulva and HSV-2 antigen was detected by direct immunofluorescence assay (IFA), HSV-2 IgG and IgM antibodies were detected by HSV-2 type specific IgG and IgM enzyme-linked immunosorbent assay (ELISA). RESULTS: HSV-2 IgG and IgM antibodies were found in 82 (63.1%) and 18 (11.3%) of 130 pregnant women. HSV-2 type-specific antigen was detected in 22 (16.9%) pregnant women by IFA test, 17 (13.1%) of whom had HSV-2 IgM antibodies. HSV-2 was isolated only in 3 women. INTERPRETATION & CONCLUSION: The seroprevalence of HSV-2 (63.1%) and genital HSV-2 infection (16.9%) was high among asymptomatic pregnant women in Adana, Turkey. Therefore, to reduce the risk of neonatal herpes, HSV-2 type-specific antibodies should be detected in pregnant women using serological tests that allow to identify women with asymptomatic or subclinical genital HSV-2 infection and those susceptible to primary genital HSV-2 infection.

Possibility of blood and hepatitis B contamination through aerosols generated during debonding procedures.

The primary purpose of this study was to investigate the presence of blood or blood elements in aerosols generated during the debonding procedures. The presence of three hepatitis B carriers in the study group led us to investigate the possibility of hepatitis B virus (HBV) contamination through aerosols, which was the secondary purpose of the study. The study group consisted of 26 patients who had a mean age of 16 +/- 2 years. Collection of aerosol samples was done using a saliva ejector that fit on the handle of the high-speed dental instrument and was attached to a mobile evacuator. A second evacuator was used to remove and collect excess fluid accumulated in the patient's mouth. The guaiac method was used to investigate the presence of occult blood in aerosol and in excess fluid samples. Serum, excess fluid, and aerosol samples of three hepatitis B carriers were tested by enzyme-linked immunosorbent assay for detecting hepatitis B surface antigen (HBsAg) and by polymerase chain reaction for detecting HBV-deoxyribonucleic acid (DNA). Blood was found in all the aerosols and in excess fluid samples. HBsAg was detected in excess fluid samples of the two hepatitis B carriers, whereas HBV-DNA was detected in only one of the excess fluid samples. HBsAg and HBV-DNA were detected in aerosol sample of only one hepatitis B carrier. The results of this study showed that aerosols generated during the debonding procedure should always be considered as potential hazards to health.

Detection of HPV DNA in cervical specimens collected in cytologic solution by ligation-dependent PCR.

OBJECTIVE: To determine the feasibility and sensitivity of detecting human papillomavirus (HPV) in specimens collected in Cytyc PreservCyt fluid (Boxborough, Massachusetts, U.S.A.) using ligation-dependent polymerase chain reaction (LD-PCR) and to demonstrate the diagnostic value of HPV DNA testing as an adjunct to cytology in the detection of cervical squamous intraepithelial lesions (SIL), especially in cases of atypical squamous cells of undetermined significance (ASCUS). STUDY DESIGN: LD-PCR is a recently invented DNA amplification technology that utilizes a capture probe for target isolation and 2 hemiprobes for target detection. The hemiprobes are designed in such a way that when they hybridize to their target, the 5' end of one probe and the 3' end of the other probe are brought together. Two hemiprobes can then be ligated into a full probe that can serve as a template for PCR amplification. A total of 94 cervical specimens were collected in cytologic fluid and tested with LD-PCR. The results were compared with those of the Digene Hybrid Capture II assay (HC II) (Beltville, Maryland, U.S.A.) and consensus PCR. RESULTS: The overall sensitivity for detecting HPV was 41.5% (39/94) by LD-PCR, 50% (47/94) by consensus PCR and 37.2% (35/94) by HC II. The prevalence of HPV by HC II, consensus PCR and LD-PCR were 87.5%, 100% and 87.5% in the high grade SIL group; 100%, 90.9% and 90.9% in the low grade SIL group; 30%, 52.5% and 40% in the ASCUS group; and 14.2%, 22.8% and 17.1% in women with normal cytology. These results indicate that all 3 methods have similar sensitivity in patients with SIL. However, there is greater variation in detection rates in the ASCUS and normal cytology groups. CONCLUSION: LD-PCR is a useful method of detecting HPV in liquid-based gynecologic cytologic preservatives, and HPV testing as a method adjunct to the liquid-based Pap test could be useful in detecting SILs, especially for the management of patients with ASCUS.