Kliničke i molekularno-epidemiološke karakteristike infekcije humanim metapneumovirusom u Hrvatskoj

Human metapneumovirus (HMPV) is an important respiratory pathogen, especially among young children. The aim of this study was to determine the incidence of HMPV infection in hospitalized children with acute respiratory tract infection (ARTI) in the season 2005/2006 in Croatia, as well as to perform the genotypic analysis of detected HMPV strains. From December 1 to March 31 2005/2006 nasopharyngeal secretions (NPSs) were collected from 402 inpatients up to 5 years of age with ARTI hospitalized in two clinics in Zagreb. NPSs were tested by real time RT-PCR assay targeting the nucleoprotein (N) gene of HMPV. To perform the phylogenetic study, partial nucleotide sequences were obtained for HMPV fusion (F) gene of 30 HMPV positive samples. HMPV was detected in 33 (8.2%), respiratory syncitial virus (RSV) in 101 (25.1%), adenoviruses in 41 (10.2%), parainfluenza viruses types1-3 in 24 patients (6.0%) and influenza viruses in 3 (0.7%) patients. The peak incidence of HMPV infection was detected in January (18/33). Three of 34 HMPV positive specimens were positive for other viral pathogens (1- parainfluenza virus type 3, 1- RSV, 1- adenovirus). No difference was noted between HMPV infected children and children infected with other viruses regarding to the sex. Children infected with adenovirus were significantly older than children infected with other viruses (HMPV, RSV and parainfluenza viruses). The majority of the infections caused by HMPV (69.7%) children presented with lower respiratory tract infections (LRTI). Comparative analysis of the clinical presentation of HMPV and RSV infected children revealed no significant differences between those two groups while adenoviruses and parainfluenza viruses caused more upper respiratory tract infections (URTI) than HMPV and RSV. Patient co-infected with parainfluenza virus type 3 and HMPV showed more severe clinical picture comparing to the clinical presentation of single-infection of HMPV in the same patient. Phylogenetic analysis showed the circulation of two main genetic lineages (A and B). It also showed the existence of two sublineages within the group B (B1 and B2) and three subclusters within lineage A (A1, A2a and A2b). The most prevalent subtype was the genotype B2 (15/30), followed by B1 (11/30), while group A of HMPV was detected in only four specimens. Interestingly, for one A2b genotype isolate epidemiological data suggests importation from Germany. All patients infected with HMPV genotype A had clinical presentation of LRTI, while 36% (9/25) of those infected with genotype B had URTI. All strains detected from patients infected with HMPV alone who required supplemental oxygen belonged to the genotype B2, while two HMPV strains from patients with co-infection who received supplemental oxygen belonged to the genotype B1. Evaluation of HMPV incidence contributes to the etiology of ARI in Croatia, while genotyping of the HMPV strains from Croatia contributes to the data of the genetic variability and molecular epidemiology of the virus in the region and in the world

Molecular mechanisms of Chlamydia trachomatis resistance to antimicrobial drugs

Chlamydia trachomatis (C. trachomatis) is a leading cause of bacterial sexually transmitted infections in developed and undeveloped countries, and therefore a global public health issue. In an era of increasing bacterial resistance to antibiotics, resistance has been an exceedingly rare phenomenon in C. trachomatis; however, clinical treatment failures attributed to multidrug-resistant C. trachomatis strains have been described on several occasions. Cell culture systems using McCoy cells and subsequent immunofluorescent staining are still the most common methodology used for antimicrobial susceptibility testing, but the presence of resistance markers should be appraised by further genetic analysis. Azithromycin resistance of C. trachomatis is often a result of the mutations in the peptidyl transferase region of 23S rRNA genes, tetracycline resistance is usually linked to the presence of foreign genomic islands integrated in chlamydial chromosome, whereas a predominant mechanism of fluoroquinolone resistance is a point mutation in the gyrA quinolone-resistance-determining region. A nucleotide substitution in rpoB gene is responsible for rifampin resistance, and different mechanisms have been involved in the development of resistance to aminoglycosides, lincomycin and sulphonamide/trimethoprim combinations

Insights on the Antimicrobial Efficacy of Azithromycin According to the First Croatian Antibiotic Susceptibility Study of Chlamydia Trachomatis Isolates from Genitourinary Tract

Bez obzira na to što je rezistencija bakterija na antibiotike (antimikrobna otpornost) jedan od glavnih izazova moderne medicine, klamidijska je infekcija dosad pokazivala dobar klinički odgovor na lijek prvog izbora – azitromicin. Ipak, u literaturi postoje ponovljeni opisi laboratorijski dokazane rezistencije spolno prenosive bakterije Chlamydia trachomatis (C. trachomatis) koja uzrokuje ovu infekciju. Osim toga, Hrvatska je zemlja s visokom stopom propisivanja azitromicina i drugih antimikrobnih lijekova, što može utjecati na pojavu otpornosti ili slabijeg odgovora na terapiju. U skladu s time cilj prvoga hrvatskog istraživanja antimikrobnog profila C. trachomatis bio je utvrditi djelotvornost azitromicina i drugih antibiotika prema urogenitalnim kliničkim izolatima C. trachomatis u laboratorijskim uvjetima. U staničnome modelu McCoyevih stanica određivane su minimalna inhibitorna koncentracija (MIC) i minimalna klamidicidna koncentracija (MCC), a svi ispitivani sojevi bili su osjetljivi na azitromicin (MIC < 4 μg/ml). Premda rezistentni sojevi bakterije nisu nađeni, kod određenih izolata uočene su znatno više vrijednosti MCC-a za azitromicin, što pokazuje trend moguće smanjene osjetljivosti. Bez obzira na zahtjevnu metodologiju, potrebno je nastaviti s ovakvim istraživanjima, poglavito u zemljama s velikim opterećenjem propisivanja antibiotika.Regardless of the fact that antimicrobial resistance represents one of the most pressing issues of modern medicine, thus far chlamydial infection responded well to the first-line therapy i.e. azithromycin. Nonetheless, laboratory-confirmed resistance of sexually-transmitted pathogen Chlamydia trachomatis (C. trachomatis) causing this infection has been repeatedly described in the literature; furthermore, Croatia belongs to the group of countries that exhibit high prescribing rates of azithromycin and other antimicrobial drugs, which can, in turn, give rise to frank resistance or decreased susceptibility. Accordingly, the first Croatian antibiotic susceptibility study of Chlamydia trachomatis aimed to assess in vitro effectiveness of azithromycin and other antibiotics against clinical chlamydial isolates from genitourinary tract. McCoy cell culture model was used to assess minimal inhibitory concentration (MIC) and minimal chlamydicidal concentration (MCC), and all the tested isolates were susceptible to azithromycin (MIC < 4 μg/ml). Although no resistant strains have been found, certain isolates showed significantly higher values of MCC for azithromycin, pointing towards the trend of possible decreased susceptibility. Regardless of the complex methodology, it is pivotal to continue with this kind of resistance surveillance, especially in countries with high antibiotic prescribing rate

Sociodemographic, sexual behaviour and microbiological profiles of men attending public health laboratory for testing for sexually transmitted diseases

In order to identify the groups at risk of sexually transmitted diseases (STDs), we assessed the sociodemographic profiles of men testing for STD, their sexual habits, and the results of microbiological analysis. During a three-year period, a total of 700 men older than 18 years of age completed the questionnaire regarding sociodemographic and sexual behavior. Urethral swabs were taken for microbiological analysis. Thirty-three percent of respondents reported not using condoms. Those that do not use condoms were predominantly less educated, unmarried but in steady relationships, employed, with children, and smokers. Alcohol or drug usage before sexual intercourse was disclosed by 21.4% of respondents, and 10.3% respondents reported sexual intercourses with commercial sex workers. Finally, 24.0% respondents reported sexual relations abroad. In 28.1% of subjects, one or more pathogens were observed in urethral swabs. The most commonly diagnosed microorganism was Ureaplasma urealyticum, followed by Chlamydia trachomatis, Mycoplasma hominis, Trichomonas vaginalis, and Neisseria gonorrhoeae. This study identified several factors that may contribute to the general risk of STD transmission, which will serve to better understand the transmission dynamics and implementation of adequate prevention programs. </p

The role of IgG avidity in diagnosis of cytomegalovirus infection in newborns and infants [Uloga IgG aviditeta u dijagnostici infekcije citomegalovirusom u novorođenčadi i dojenčadi]

To evaluate the value of IgG avidity in diagnosis of congenital cytomegalovirus (CMV) infection in newborns and infants we collected serum samples from 40 infants under 12 months of age with suspected congenital CMV infection. Sera were tested for IgM, IgG and IgG avidity. For 25 of them, virus isolation and/or polymerase chain reaction (PCR) on urine specimens were performed. Thirteen (32.5%) patients showed the presence of CMV IgM antibodies, 3 (7.5%) had equivocal IgM result, and 24 (60.0%) patients had IgG antibodies only. Using IgG avidity, CMV infection (low avidity index-AI) was documented in 61.5% IgM positive and 54.2% IgM negative patients. Eight of nine (88.8%) IgM positive patients were positive either on virus isolation or PCR. In IgM negative patients, 46.6% urine cultures were positive for CMV and 66.6% were PCR positive. According to age, IgG avidity demonstrated acute/recent primary CMV infection in 58.8% patients younger than three months compared with 91.7% and 81.8% in 3-6 and 6-12 months old babies, respectively. In conclusion, IgG avidity is useful in diagnosis of CMV infection either in IgM positive or IgM negative children older than 3 months of age. In infants less than 3 months, transplacentally derived maternal IgG antibodies of high avidity influence on the IgG avidity result. In these children, CMV infection should be confirmed by direct virologic methods such as virus isolation or PCR

Značaj IgG aviditeta u dijagnostici infekcije citomegalovirusom u trudnica i novorođenčadi

Aim: To asses the value of IgG avidity in diagnosis of CMV infection in pregnant women and newborn infants. Methods: During the three-year period (2003-2005) serum samples from 64 pregnant women and 32 infants less than 12 months of age with suspected congenital/perinatal CMV infection were collected. Sera were tested for CMV IgM and IgG using an indirect enzyme-linked immunosorbent assay and IgG avidity using urea as denaturing agent. Results: Among IgM positive women, 2/26 (7.6%) showed an increase of IgG avidity index (AI) from intermediate to high AI in paired sera samples indicating recent primary infection and 24/26 (92.4%) showed high AI indicating past infection. All women with negative IgM antibodies had high AI. In infants less than 12 months old, acute/recent primary CMV infection was documented in 8/12 (66.7%) children with positive IgM and in 10/20 (50.0%) children with negative IgM antibodies. In two children less than three months with high AI, CMV infection was confirmed by virus isolation from urine. One of them showed a decrease of AI from high to low in paired sera samples. Conclusions: IgG avidity differentiates primary from nonprimary CMV infection in both pregnant women and infants older than three months. In children less than 3 months of age, transplacentally transferred maternal antibodies of high avidity may influence on the serologic test results.Cilj rada: Ispitati vrijednost IgG aviditeta u dijagnostici CMV infekcije u trudnica i novorodencadi. Metode: Tijekom trogodišnjeg perioda (2003-2005) prikupljeni su uzorci seruma od 64 trudnice, te 32 djece do godinu dana starosti sa suspektnom kongenitalnom/perinatalnom CMV infekcijom. Serumi su testirani na prisustvo CMV IgM i IgG protutijela metodom ELISA, te aviditet IgG protutijela uz primjenu uree. Rezultati: U skupini trudnica s pozitivnim IgM protutijelima, u njih 2/26 (7,6%) dokazan je porast indeksa IgG aviditeta (AI), od granicnog u visoki AI u parnim uzorcima seruma, što ukazuje na nedavnu primarnu CMV infekciju, dok je 24/26 (92,4%) imalo visok AI (prošla CMV infekcija). Sve trudnice s negativnim IgM protutijelima imale su visok AI. U djece mlade od 12 mjeseci, akutna/nedavna primarna CMV infekcija dokazana je u 8/12 (66,7%) djece s pozitivnim IgM, te u 10/20 (50,0%) djece s negativnim IgM protutijelima. U dvoje djece mlade od 3 mjeseca s visokim AI, CMV infekcija je potvrdena izolacijom virusa iz urina. U jednog od njih došlo je do pada AI iz visokog u niski, u parnom uzorku seruma. Zakljucci: Pomocu testa IgG aviditeta moguce je razluciti primarnu od prošle CMV infekcije u trudnica i djece starije od 3 mjeseca. U djece mlade od 3 mjeseca, transplacentarno prenesena majcina IgG protutijela visokog aviditeta mogu utjecati na rezultate seroloških pretraga

Mycoplasma genitalium – sexually transmitted pathogen that cannot be ignored

Iako je razvoj molekularnih metoda omogućio učinkovitu dijagnostiku infekcije uzrokovane bakterijom Mycoplasma genitalium (M. genitalium), podaci o infekcijama uzrokovanih ovom bakterijom u Hrvatskoj su nedostatni, budući je dijagnostika vrlo zahtjevna te još uvijek nije rutinska metoda u mikrobiološkim laboratorijima. Cilj ovog rada bio je utvrditi učestalost M. genitalium u obriscima cerviksa i obriscima uretre muškaraca zaprimljenih u Službi za kliničku mikrobiologiju Nastavnog zavoda za javno zdravstvo "Dr. Andrija Štampar" radi testiranja na C. trachomatis. Istraživanje je provedeno u razdoblju od sedam mjeseci. Od ukupno 5057 zaprimljenih obrisaka cerviksa koji su bili testirani na C. trachomatis, 195 uzoraka je testirano na M. genitalium (99 C. trachomatis pozitivnih i 96 C. trachomatis negativnih). Testirano je i 52 obrisaka uretre muškaraca s negonokoknim uretritisom (18 C. trachomatis pozitivnih i 34 C. trachomatis negativnih). M. genitalium i C. trachomatis dokazivane su real-time PCR metodama, dok je gonokokna infekcija isključena pregledom izravnog mikroskopskog preparata. U obriscima cerviksa učestalost infekcije uzrokovane C. trachomatis iznosila je 2,04% (103/5057). Od 195 testiranih obrisaka cerviksa, M. genitalium je dokazana u 6 uzoraka (3,08%). Svi obrisci cerviksa pozitivni na M. genitalium bili su istodobno pozitivni i na C. trachomatis (6/99) dok niti jedan C. trachomatis negativan uzorak nije bio pozitivan na M. genitalium (0/96; P < 0,05). U obriscima uretre muškaraca s negonokoknim uretritisom M. genitalium je dokazana u jednom uzorku koji je bio C. trachomatis negativan. M. genitalium je važan, ali u našoj sredini nedovoljno prepoznat i nedovoljno dijagnosticiran uzročnik spolno prenosivih bolesti. Sukladno rezultatima ove studije testiranje na M. genitalium opravdano je izvoditi u sklopu rutinskog testiranja na spolno prenosive bolesti u obriscima cerviksa žena kod kojih je dokazana C. trachomatis infekcija.The development of molecular methods has enabled efficient diagnosis of infections caused by Mycoplasma genitalium (M. genitalium). However, data on M. genitalium infection in Croatia are scarce due to the complexity of detection method for this bacterium which is still not routinely used in clinical laboratories. The aim of this study was to determine the prevalence of M. genitalium in cervical swabs and male urethral swabs referred for C. trachomatis diagnostics to the Clinical Microbiology Department of the Teaching Institute for Public Health "Dr. Andrija Štampar". The study was conducted over a period of seven months. From a total of 5057 received cervical swabs that were tested for C. trachomatis, 195 samples were tested for M. genitalium (99 positive and 96 negative for C. trachomatis). Additionally, 52 urethral swabs collected from men with nongonococcal urethritis were tested for M. genitalium (18 positive and 34 negative for C. trachomatis). Both, C. trachomatis and M. genitalium were detected using realtime PCR methods, while gonococcal infection was excluded by direct microscopy. The incidence of C. trachomatis infection in cervical swabs was 2.04% (103/5057). Out of 195 cervical swabs tested M. genitalium was proved in six samples (3.08%). All samples found positive for M. genitalium were positive for C. trachomatis as well (6/99), while not a single C. trachomatis negative sample was positive for M. genitalium (0/96; P < 0.05). In the urethral swabs of men with NGU, M. genitalium was detected in one sample that was C. trachomatis negative. M. genitalium is important, but in our midst under-recognized and under-diagnosed sexually transmitted pathogen. Based on the results of the study, testing for M. genitalium should become an integral part of routine protocol for diagnosis of sexually transmitted diseases in women with C. trachomatis infection

WEST NILE VIRUS INFECTION: RE-EMERGENT DISEASE IN CROATIA

Virus Zapadnog Nila (VZN) malen je, ovijen, kuglast virus koji pripada porodici Flaviviridae, roduFlavivirus, serokompleksu japanskog encefalitisa. Prirodni rezervoari VZN-a su ptice, a glavni vektori komarci roda Culex. Do sada je opisano sedam genskih linija VZN-a, od kojih su najrasprostranjenije linije 1 i 2 (Afrika, Sjeverna i Južna Amerika, Europa, Azija i Australija). Infekcija VZN-om u oko 80% slučajeva prolazi asimptomatski. U 20% zaraženih javlja se nespecifična febrilna bolest (VZN-groznica), a u manje od 1% razvija se neuroinvazivni oblik bolesti (meningitis, encefalitis, sindrom poliomijelitisa). Na području Hrvatske protutijela na VZN dokazana su u ljudi, medvjeda i konja. U kolovozu i rujnu 2012. godine prvi put su zabilježeni klinički slučajevi neuroinvazivne infekcije VZN-om u ljudi te asimptomatska akutna infekcija u konja u tri županije istočne Hrvatske. Dijagnoza je potvrđena serološkim testovima (imunoenzimni test, IgG-aviditet, neutralizacijski test redukcije plakova).West Nile virus (WNV) is a small, enveloped, spherical virus that belongs to the family Flaviviridae, genus Flavivirus, Japanese encephalitis serocomplex. Natural reservoirs of WNV are birds, and the main vectors are mosquitoes of the genus Culex. There are seven genetic lineages of WNV. Lineages 1 and 2 are the most widely distributed (Africa, North and South America, Europe, Asia and Australia). About 80% of infections are asymptomatic. In 20% of patients nonspecific febrile disease occurs (West Nile fever). Less than 1% of infected persons will develop neuroinvasive WNV disease (meningitis, encephalitis, and poliomyelitis). In Croatia, antibodies to WNV were demonstrated in humans, bears and horses. In August-September 2012 clinical cases of human WNV neuroinvasive disease and asymptomatic acute infection in horses were reported for the first time in three eastern Croatian counties. The diagnosis was confirmed by serologic tests (enzyme immunoassay, IgG avidity, plaque-reduction neutralization test)

DIE HAUFIGKEIT POSTTRAUMATISCHER STRESSREAKTIONEN UNTER VERTRIEBENEN UND DER MOGLICHE BEZUG ZUR INHAFTIERUNG IN GEFANGENENLAGERN

Cilj rada bio je utvrditi učestalost PTSP-a u prognanika iz Hrvatske te ispitati povezanost PTSP-a s brojem trauma, posebno s boravkom u logoru, te s demografskim varijablama. Ispitana je grupa od 88 prognanika, od kojih 34 muškarca i 54 žene, prosječne dobi 43 godine. Primijenjeni su upitnik za traume i Watsonov intervju za PTSP. Grupa je proživjela prosječno tri traume, a učestalost PTSP-a nakon dvije godine života u progonstvu iznosila je 45,45%. Ustanovljeno je da se pojavnost ukupne izraženosti simptomatike PTSP-a povećava s povećanjem broja traumatskih događaja i s dobi osobe, a smanjuje se s većim obrazovanjem osobe. Grupa prognanika (N=18) koji su bili zatočeni u logoru do najviše mjesec dana, a koji nisu bili izloženi torturi, uspoređena je s komparativnom grupom prognanika (N=18) izjednačenom po dobi, spolu, obrazovanju i broju proživljenih trauma. Grupe se nisu statistički značajno razlikovale u izraženosti ukupne simptomatike PTSP-a. Zaključno, rezultati ukazuju da su mlađa odrasla dob i obrazovanje protektivni čimbenici u reakciji na traumu. Kratko zatočeništvo u logoru, bez torture, nije imalo zaseban efekt na pojavnost simptomatike PTSP-a u politraumatiziranoj grupi prognanika.The aim of the study was to explore the prevalence of PTSD in refugees from Croatia and investigate the relationship between PTSD and the number of experienced traumas, especially with regard to being held in detention camp, as well as the relationship between PTSD and demographic variables. A group of refugees, consisting of 34 men and 54 women, mean age 43, were studied. Trauma Questionnaire and Watson\u27s PTSD Interview were applied. The group experienced three traumas, on average; whereas after two years of life in captivity the prevalence of PTSD was 45.45%. It was established that the total of PTSD symptoms grow with the increasing number of traumas and with the refugee\u27s age, but decrease with the person\u27s higher education. The group of refugees (N=18) who were kept in detention camp no longer than one month and were not tortured, was compared with the comparative refugee group (N=18), equal in gender, age, education and number of traumas experienced. The groups had shown statistically the same amount of total PTSD symptomatics. Conclusively, the results indicate that young adulthood and higher education are protective factors in reaction to trauma. A short period of time spent in detention camp, without torture, did not have a separate effect on PTSD symptomatics in the group of po- Iytraumatized adult refugees.Die vorliegende Arbeit soll ermitteln, wie haufig posttraumatische Stressreaktionen unter kroatischen Vertriebenen auftreten und inwiefern diese Stressreaktionen mit der Zahl der durch- Iebten Traumata, insbesondere mit der Inhaftierung in Gefangenenlagern sowie mit demographischen Variablen in Verbindung stehen. Die befragte Personengruppe umfasste 88 Vertriebene: 34 Manner und 54 Frauen im Durchschnittsalter von 43 Jahren. Angewandt wurden ein Fragebagen zur Ermittlung erlebter Traumata sowie der Watsonsche Fragebagen zur Ermittlung posttra umatischer Stressreaktionen. Die befragten Personen erlebten im Durchschnitt jeweils 3 Traumata, die Haufigkeit posttraumatischer Stressreaktionen zwei Jahre nach der Vertreibung betrug 45,45%. Es wurde festgestelIt, dass sich mit der Zahl traumatischer Erlebnisse und mit dem wachsenden Alter der Befragten die Gesamtauspraqunq des posttraumatischen Stresssyndroms steigert, mit einem htiheren Bildungsgrad jedoch abfallt, Eine Gruppe von Vertriebenen (N=18), die bis zu 1 Manat in Lagern inhaftiert gewesen, jeda ch keinen Folterungen ausgesetzt waren, wurde mit einer Vergleichsgruppe von Vertriebenen (N=18) kompariert, mit der sie in Alter, Geschlecht, Bildungsgrad und der Zahl durchlebter Traumata Obereinstimmte. Die Gruppen unterschieden sich statistisch nur wenig in der Gesamtauspragung posttraumatischer Stressreaktionen. Es kann gemar.. den Resuitaten abschliefšend gesagt werden, dass ein jOngeres Erwachsenenalter und ein h6herer Bildungsgrad Schutzfaktoren bei der Reaktion auf Traumata darstellen. Kurze Lageraufenthalte ohne Folterungen hatten keinerlei Sonderwirkung auf die Haufigkeit posttraumatischer Stressreaktionen unter den befragten polytraumatisierten Vertriebenen