COVID-19 autopsies of Istanbul.

BACKGROUND/AIMS: The aim of this study is to share autopsy findings of COVID-19-positive cases and autopsy algorithms for safely handling of suspicious bodies during this pandemic. METHODS: COVID-19-positive cases of Istanbul Morgue Department were retrospectively analyzed. Sampling indications for PCR tests in suspicious deaths, macroscopic and microscopic findings obtained in cases with positive PCR tests were evaluated. RESULTS: In the morgue department, 345(25.8%) of overall 1336 autopsy cases were tested for COVID-19. PCR test was found positive in 26 cases. Limited autopsy procedure was performed in 7 cases, while the cause of death was determined by external examination in the remaining 19 cases. Male-to-female ratio was found 3.3:1 and mean age was 60.0 ± 13.6 among all PCR-positive cases. Cause of death was determined as viral pneumonia in fully autopsied cases. Most common findings were sticky gelatinous fluid in cavities and firm and swollen lungs, varying degrees of consolidation. In microscopy, diffuse alveolar epithelial damage, type-II pneumocyte hyperplasia, hyaline membrane formation, fibrinous exudate, and fibrinous plaques in the alveoli were the most common findings. CONCLUSIONS: In COVID-19 autopsies, pulmonary findings were found to be prominent and the main pathology was pneumonia. Older age and findings of chronic diseases indicate that the cases were in the multirisk group in terms of COVID-19 mortality.

[Postmortem Molecular Epidemiology of HIV-1 Strains Isolated in Turkey]. / Türkiye'de Postmortem Izole Edilen HIV-1 Suslarinda Moleküler Epidemiyoloji.

Human immunodeficiency virus (HIV) comprises two genotypes, namely HIV-1 (group M, N, O and P) and HIV-2 (group A to H), which differ in their envelope glycoproteins and other antigenic epitopes despite their morphological and biological resemblance.Group M of HIV-1 responsible for 95% of HIV infections worldwide is composed of nine subgroups. In addition to subgroups, group M contains also two recombinant forms, known as circulating recombinant form (CRF) and unique recombinant form (URF). The first case of HIV/acquired immun deficiency virus (AIDS) in Turkey was reported in 1985 and the current number of cases reached a total of 18.557 including 1736 with AIDS based upon the surveillance data of Ministry of Health between October 1985 and November 2018. The aim of this study was to determine the prevalence of HIV-1 strains isolated from HIV positive autopsy cases detected by HIV polymerase chain reaction (PCR) and determine drug resistance. Twenty eight cases [17 males, 11 female: age ranged between 3 months and 66 years (median: 35 years)] found to be HIV positive among the autopsy cases sent for HIV1 PCR study and serological screening between 2011-2017 were recruited in the study. For identification of subtypes in HIV-1 isolates, most-preferred analysis tool was used [HIVdb Stanford University Genotypic Resistance Interpretation Algorithm (www.hivdb.stanford.edu)]. Phylogenetic tree was made according to direct sequencing of HIV-1 reverse transcriptase (pol) region and phylogenetic analysis was evaluated in 23 cases. Los Alamos National Laboratory were trimmed from full-length genomes. Phylogenetic analysis of the 870 base pair of the pol gene region was performed using CLC Sequence Viewer v8.0 (Qiagen Aarhus A/S, www.qiagenbioinformatics.com) software. The phylogenetic tree was obtained according to the neighbor-joining method and the Jukes-Cantor nucleotide distance scale and bootstrap value was set at 1000. In our study, subtype B was found to be most frequent type (39.3%; 11/28). Subtype A (17.9%; 5/28), CRF02_AG (14.3%; 4/28), subtype C (10.7%; 3/28), B+CRF02_AG recombinant (3.6%; 1/28), CRF01_AE (3.6%; 1/28), subtype D (3.6%; 1/28), as well as subtype F (3.6%; 1/28) and subtype G (3.6%; 1/28) strains were also detected in the circulation. Analysis of our results showed that 32.1% (9/28) of the samples exhibited resistance mutations. Detected mutations were as follows: M41L, T215C, K65R, M184V, responsible for nucleoside reverse transcriptase inhibitor (NRTI) resistance; K103N, Y181C, G190A, responsible for non-nucleoside reverse transcriptase inhibitor (NNRTI) resistance; D30N, M46I, responsible for protease inhibitor (PI) resistance. NRTI, NNRTI and PI mutation rates in the samples were found as 21.4%, 7.1% and 3.6%, respectively. Although number of samples analyzed in our study is low, we can propose that they resemble the strains circulating in Turkey. The results of our study; although the subtype B is still dominant in our country, it supports other studies reporting that there are non-B subtypes and an increase in CRF rates in recent years. Phylogenetic analysis is widely regarded as the gold standard technique to determine the subtypes of HIV-1. Molecular epidemiologic studies related to HIV may be important in monitoring HIV subtype patterns and spreading pathways in that country. As a result; the opportunity to collect postmortem HIV sequences in a database appears to have occurred, and as this database expands, its usability is available. Therefore, it is thought that HIV subtypes and mutation information may be useful.

[Investigation of viral respiratory tract infection agents by multiplex PCR method in autopsy cases: A five-year study]. / Otopsi olgularinda viral solunum yolu enfeksiyonu etkenlerinin multipleks PCR yöntemi ile arastirilmasi: bes yillik bir çalisma.

Viral respiratory infections are one of the leading causes of morbidity and mortality, especially in children, elderly and immunocompromised patients. The inclusion of post-mortem studies to diagnose the infection causing mortality could be beneficial in specifying new pathogens and determining strategies for treatment and prevention. The aim of this study was to research viral etiology by applying multiplex real-time polymerase chain reaction (Rt-PCR) method in autopsy cases who have been considered to have a respiratory infection and to assess whether the viruses detected are the primary cause of the infection and whether they have any contributory effect on the mortality together with histopathological evidence. In this study, we included a total of 834 cases consisting of sudden death cases from infantile-pediatric age group and autopsy cases from > 18 year age group with suspected respiratory tract infection in our laboratory between January 2013 and May 2017. Of 834 cases, 468 (56.1%) were male and 366 (43.9%) were female, there were 191 (22.9%) cases between 0-1 months, 593 (71.1%) cases between 1 month-18 years, and 50 (6%) cases in the > 18 years age group. In 728 of 834 (87.3%) cases nasopharyngeal/tracheal swab samples and in 106 (12.7%) of them paraffin-embedded lung tissue samples were studied by the use of "FTD Respiratory 21 (Fast-Tract Diagnostics Luxemburg)" kit, with multiplex Rt-PCR method. The post-mortem samples were evaluated for human rhinovirus (HRV), parainfluenza viruses (PIV) (1, 2, 3, 4), influenza virus type A and B (INF-A, INF-B), enterovirus (EV), human bocavirus (HBoV), adenovirus (AdV), human coronavirus (HCoV 229,63,HKU,43), human metapneumovirus A ve B (HMPV-A/B), parechovirus, respiratory syncytial virus (RSV A/B) and Mycoplasma pneumoniae. In our study, at least one respiratory virus was detected by Rt-PCR in 379 (45.4%) of total 834 cases, whereas no viral agent was identified in 455 (54.6%) of the cases. One viral agent was detected in 278 (33.3%), two viral agents were detected in 83 (9.94%) and three viral agents were detected in 18 (2.16%) cases. Overall, the most common viral agent was HRV 110 (13.2%) followed by AdV 39 (4.7%) and RSV A/B 33 (4%). In pediatric cases the rate of positive results for respiratory viruses was 31.8% and in adult group it was 20% (p= 0.032). The most common virus detected among children was HRV and INF-A in adult group. In 101 (12.1%) cases infections caused by two or three agents were diagnosed. Infections with two causative agents were detected as 2.6% (5/191) in 0-1 month age group, 13% (77/593) in 1 month-18 year age group and 2% (1/50) in > 18 age group. The most frequently observed co-infections with double causative agents were HRV and INF-B, HRV and PIV, HRV and HBoV, HRV and AdV combinations. Infections with three causative agents were detected completely among 1 month-18 year age [3% (18/593)] group. In our study, 318 (38.1%) cases had no signs of infection in the postmortem histopathological examination of the lung tissues, while the most common finding was lobular pneumonia/purulent bronchitis in 233 (28%) cases and the second was interstitial pneumonia in 168 (20.1%) cases. When all cases were evaluated in terms of infection, positive results were detected in 469 (56.2%) cases. As a result; postmortem microbiological diagnosis with autopsy and histopathological detection of the patients who are thought to have respiratory tract infection will also determine the infectious agents causing death.

Evaluation of Virulence Factors and Antifungal Susceptibility in Yeast Isolates from Postmortem Specimens.

Invasive fungal infections are a leading cause of morbidity and mortality in immunocompromised patients, especially in cases requiring a prolonged stay in the intensive care unit. A total of 99 yeast strains were isolated from 42 postmortem cases. In this study, virulence factors and antifungal susceptibility of these species were evaluated. The isolates were identified as Candida albicans (54), C. tropicalis (15), C. glabrata (12), C. parapsilosis (6), C. lipolytica (3), C. utilis (3), C. krusei (2), C. kefyr (1), and Cryptococcus neoformans (3). The most commonly isolated species was C. albicans, and no resistant species were determined. Despite the equal number of specimens, no secretion of significant virulence factors was associated with the postmortem specimen in the Candida species. Postmortem fungal investigations in forensic autopsies are useful in explaining cause of death in such cases, also may lead to protocols for the treatment of fungal infections and contribute to fungal pathogenesis and epidemiological data.

Postmortem diagnosis of cytomegalovirus and accompanying other infection agents by real-time PCR in cases of sudden unexpected death in infancy (SUDI).

As an opportunistic pathogen with high mortality rates, Cytomegalovirus (CMV) may lead to fatal disseminated CMV infection of the premature and newborn; thus necessitating the demonstration of CMV-DNA with clinical history and/or histopathological findings of CMV infection and defining other bacterial and viral infection agents with real-time polymerase chain reaction (RT-PCR) in udden unexpected death in infancy (SUDI) cases as we aimed in this study. 314 (144 female, 170 male) SUDI cases were prospectively investigated from January 2013 to January 2015 in Istanbul Forensic Medicine Institution. The study includes 87 tissue samples of 39 cases for post-mortem histopathological examination of interstitial pneumonia, myocarditis, meningitis, encephalitis, hepatitis, colitis or tubulointerstitial nephritis and/or accompanying chronic sialadenitis. CMV-DNA was found positive in 35 (40.2%) salivary gland, 19 (21.8%) lung, 1 (1.1%) tonsil, and 1 (1.1%) brain tissues. CMV sialadenitis and/or CMV pneumonia associated with other viral and/or bacterial agents were detected in 23 (60%) of 39 infant cases. The demonstration of CMV-DNA would significantly clarify the cause of death and collection of epidemiological data in SUDI cases with clinical history and histopathological findings of CMV infection accompanying chronic CMV sialadenitis. Furthermore, CMV suppresses the immune system, and may predispose to other bacterial and/or viral infections in these cases. Post-mortem molecular investigations are useful in explaining cause of death in SUDI with a suspicion of infection in forensic autopsies.

Detection of Human Bocavirus DNA by Multiplex PCR Analysis: Postmortem Case Report.

BACKGROUND: Human bocavirus (HBoV) is a virus belonging to the Parvoviridae family, which has been newly discovered to be associated with respiratory tract infections in children. There are many reports worldwide on the endemicity of this virus. Since it is relatively new, it is not routinely detected in clinical laboratory investigations. CASE REPORT: We demonstrated that HBoV infection caused the death of a 5-month-old girl with a history of high fever and wheezing. Human bocavirus (HBoV 1/2/3/4) was found in a nasopharyngeal swab, paraffin-embedded lung tissue and stool samples by multiplex PCR methods using postmortem microbiological analysis. CONCLUSION: This case suggests that lower respiratory tract infections due to HBoV may cause severe and life-threatening diseases. Postmortem microbiology is useful in both clinical and forensic autopsies, and allows a suspected infection to be confirmed. To our knowledge, this report is the first document of a HBoV postmortem case in Turkey.

[Trends in antibiotic susceptibility patterns of Streptococcus pneumoniae and Haemophilus influenzae isolates: four years follow up]. / Streptococcus pneumoniae ve Haemophilus influenzae izolatlarinin antibiyotik duyarlilik paternlerindeki degisim: dört yillik izlem.

This study was aimed to follow up the antibiotic resistance trends in Streptococcus pneumoniae and Haemophilus influenzae isolated from clinical specimens between 2003-2006 at Marmara University Hospital, Istanbul, Turkey. Antibiotic susceptibilities were performed by disk diffusion method, and penicillin susceptibility was determined by E-test (AB Biodisk, Sweden). Results were evaluated by CLSI standards. During this period a total of 258 S. pneumoniae and 548 H. influenzae were isolated in our laboratory. According to the 2006 CLSI penicillin breakpoints, overall resistance of S. pneumoniae isolates to penicillin was 39.9% and intermediate and high level penicillin resistance rates were 30.2% and 9.7%, respectively. The rates of high level penicillin resistant pneumococci by years were 11.1% in 2003; 10.9% in 2004; 6% in 2005, 12.1% in 2006 and except for 2005 no significant change in resistance rates was detected. However, according to the 2008 CLSI penicillin breakpoints, resistance was found to be 3.5%, intermediate and high level penicillin resistance being 3.1% and 0.4%, respectively. While penicillin resistance rates by years were as 4.4% in 2003, 5.5% in 2004, 0% in 2005 and 4.4% in 2006, high level penicillin resistance was detected only in 2003 as 2.2%. Resistance rates of chloramphenicol, erythromycin, tetracyline and trimethoprim-sulphametoxazole (TMP-SMX) were detected as 10.1%, 19%, 26.8% and 49.2%, respectively while erythromycin, tetracycline and TMP-SMX multi-drug resistance was detected in 12.4% of the isolates. No resistance was detected to vancomycin. Beta-lactamase production rate in H. influenzae isolates was 3.3%, being 1.6% in 2003 with a raise up to 4% in 2006. No beta-lactamase negative ampicillin-resistant isolate was detected. Although chloramphenicol and cefaclor resistance were in low levels (2.2% and 0.7%, respectively), TMP-SMX resistance was detected as 25.5%. TMP-SMX resistance was two fold more in beta-lactamase producers compared with the non-producers, whereas chloramphenicol resistance revealed a significant increase in beta-lactamase producers (1% versus 44.5%). In conclusion, doubling of beta-lactamase production rate in H. influenzae within years indicates the importance of continuous follow-up of antibiotic resistance in specific pathogens. The evaluation of penicillin results obtained for pneumococci according to modified 2008 CLSI criteria revealed that penicillin can still be used effectively in the treatment of pneumococcal respiratory tract infections. Continuous active surveillance of resistance rates provides important data for the determination of the empirical therapy protocols for S. pneumoniae and H. influenzae infections.