hossein mirhendi

SARS-CoV-2 disease is a highly contagious infection causing a large number of deaths in susceptible individuals throughout the world. In this study, a low-cost, sensitive, and easy-to-perform conventional polymerase chain reaction (PCR)-based RNA detection method was evaluated to diagnose the infection, which was feasible at a laboratory with minimal molecular infrastructure.

From 4 July to 31 August 2020, a total of 277 nasopharyngeal/oropharyngeal swab samples consisting of 72 samples from hospitalized patients with a severe respiratory infection and 205 suspected patients in Isfahan, Iran, were tested using probe-based rtRT-PCR and conventional PCR assays.

A total of 160 clinical samples were tested by rtRT-PCR using the E gene. The sensitivity and specificity of the conventional PCR method were determined to be 100%. Furthermore, out of 117 clinical samples evaluated by the probe-based RT-PCR using the N gene, 74.4% of the samples were positive. Moreover, the duplex PCR method using the N gene and RNase P as an internal control reference gene showed that 68.4% of the samples were positive. Therefore, the tested PCRs could detect positive samples with a sensitivity of 92.55% and a specificity of 100%.

According to the results, this method is a simple, inexpensive, and valuable alternative as well as a suitable procedure for the laboratory diagnosis of SARS-CoV-2 infection.

Candida auris, a multidrug-resistant yeast, can cause primary or secondary infections in a wide range of patients, including those diagnosed with the new coronavirus to even healthy individuals. The fungus has been reported in less than a decade on all six continents and in more than 45 countries. Ease of distribution, long shelf life, and resistance to several antifungal drugs have raised concerns about the prevention and management of patients with C. auris infection. Recent reports indicate serious challenges in identifying, understanding the mechanism of drug resistance, and preventing mortality from the infection with this microorganism. Given the prevalence of COVID-19 infection, it is important to identify patients colonized with C. auris correctly and at the early stages, to control and prevent a possible outbreak. In this article, the widespread occurrence of infections due to C. auris in the world and Iran, its clinical manifestations, risk factors, pathogenic mechanisms, diagnostic enhancements and challenges, drug resistance, treatment options, prevention, and control as well as concomitant C. auris infections in patients with COVID-19 virus, are reviewed.

The multidrug-resistant fungal pathogen Candida auris has been associated with healthcare. We need to address COVID-19 pandemic as well as the ongoing global fungal epidemic caused by C. auris, a multi-drug-resistant fungus spreading rapidly throughout the world. This study was conducted on patients admitted to an ICU in Isfahan, Iran, from November 2020 to February 2021 to determine the spectrum of C. auris colonization in immunocompromised patients hospitalized in an intensive care unit (ICU). Therefore, clinical swabs were collected from 32 immunocompromised patients for C. auris detection upon ICU admission after 7 to 14 days. A rich culture medium was used to evaluate C. auris growth at a higher temperature (40°C) and salinity (10% wt/vol) in Sabouraud dextrose agar, which can be used in combination with a C. auris-specific polymerase chain reaction method. C. auris was not isolated in the clinical samples of patients. The most common colonizer was Candida albicans, followed by C. glabrata, C. parapsilosis, and C. tropicalis. Candida glabrata was the only species with noticeable growth in the Salt SDA, with D-Mannitol as a carbon source. Currently, C. auris is not a common cause of systemic or superficial fungal infections in Iran. The screening of patients admitted to the ICU for C. auris could aid in the identification of colonized patients and could simplify the application of infection control measures.

Since December 2019, the world is struggling with an outbreak of coronavirus disease?2019 (COVID?19) infection mostly represented as an acute respiratory distress syndrome and has turned into the most critical health issue worldwide. Limited information is available about the association between dynamic changes in the naso/oropharyngeal viral shedding in infected patients and biomarkers, aiming to be assessed in the current study.

This quasi?cohort study was conducted on 31 patients with moderate severity of COVID?19  anifestations, whose real?time polymerase chain reaction (RT?PCR) test waspositive for severe acute respiratory syndrome?coronavirus?2 (SARS?CoV?2) RNA at baseline. RT?PCR was rechecked for patients every 3–4 days until achieving two negative ones. In parallel, biomarkers, including lymphocyte count, lactate dehydrogenase (LDH), and C?reactive protein (CRP), were assessed every other day, as well. Viral shedding also was assessed.

Spearman’s correlation test revealed a significant direct correlation between the viral shedding from the symptom onset and the time, in whichCRP (P = 0.0015, r = 0.54) and LDH (P = 0.001, r = 0.6207) return to normal levels after symptom onset, but not for lymphocyte count (P = 0.068, r = 0.34).

Based on the current study’s findings, the duration of SARS?CoV?2 RNA shedding was directly correlated with the required time for LDH and CRP return to normal levels. Therefore, these factors can be considered the determinants for patients’ discharge, isolation, and return to social activities; however, further investigations are required to generalize the outcomes.

Early and cost‑effective diagnosis and monitoring of the infection caused by severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) are critically important to anticipate and control the disease. We aimed to set up a SYBR Green‑based one‑step real‑time polymerase chain reaction (PCR) as a lower‑cost alternative method to detect the virus.

An in‑house SYBR Green‑based PCR assay targeting the envelope (E) and RNA‑dependent RNA polymerase (RdRp) genes, was set up to diagnose the infection, and was compared with the reference probe‑based PCR method.

When the commercial probe‑based assay was considered as the reference method, SYBR Green‑based PCR had a slightly lower sensitivity (81.98% and 86.25% for E and RdRp targets, respectively) and a good specificity (100% and 94.44% for E and RdRp targets, respectively). For both gene targets, three different melting temperature (Tm) patterns were found in the PCRs of the nasopharyngeal/oropharyngeal swab samples, but no size polymorphism was seen in agarose gel electrophoresis.

Further studies to improvement of the assay are needed to make it an inexpensive and reliable tool for the diagnosis of COVID‑19.

Given the important role of Malassezia spp. in skin diseasesand other associated infections in neonates, this study aimed to investigate the presenceand frequency of Malassezia spp. in the skin of neonates hospitalized in neonatalintensive care units and their mothers using culture and accurate molecular-basedmethods.

In total, 205 samples were collected from 130 neonates (>4-day-old) and 75 mothers. Isolation of Malassezia spp. from the skin was performed usingLeeming-Notman agar and modified Dixon agar media. To compare the Malasseziamicroflora on the skin of the neonates and their mothers, a polymerase chain reactionsequencing method was performed for spp. identification of 92 isolates obtained fromneonates and their mothers. Moreover, possible associated risk factors for thecolonization of Malassezia spp. on the skin were recorded.

Cultures from 62.3% of neonates and 77.3% of mothers were positive forMalassezia spp. growth. Malassezia globosa was the most prevalent isolated spp. foundin the skin of the study population. It is noteworthy that a rare Malassezia spp.,Malassezia arunalokei, was isolated from the skin of one neonate. There was a 76%similarity between the mother-neonate isolate sequences results. The statistical analysisshowed that the type of feeding is a significant (P<0.001) associated factor forMalassezia skin colonization.

The findings support the hypothesis that the colonization of Malassezia inneonates is significantly influenced by that of the mother, and this may be associatedwith breastfeeding.

The frequency and genetic diversity of black fungi in environmental and clinical settings have not been fully studied in Iran. This study aimed to identify and evaluate intra- and inter-species DNA sequence variation and also understand the phylogenetic relationships of melanized fungi and relatives isolated from different geographical regions of Iran.

In total, 111 clinical and environmental strains of dematiaceous fungi were isolated, and their internal transcribed spacer ribosomal DNA(rDNA) regions were sequenced and analyzed.

An inter-species nucleotide sequence diversity rate of 1 to 464 nucleotides was observed between the species. Intra-species differences were found in the strains of Alternaria alternata, Cladosporium cladosporioides, Alternaria tenuissima, Curvularia spicifera, Aureobasidium pullulans, Curvularia hawaiiensis, Neoscytalidium dimidiatum,Alternaria terricola, Alternaria chlamydospora, Didymella glomerata, and Drechslera dematioidea by 0–59, 0–22, 0–4, 0–4, 0–3, 0–2, 0–2, 0–2, 0–2, 0–1, and 0–1 nt, respectively.

The internal transcribed spacer rDNA is useful for the discrimination of several taxa of dematiaceous fungi. However, a better understanding of the taxonomy of species of Alternaria requires a larger rDNA region or a library of other gene sequences.

The taxonomy of Candida is controversial and has undergone changes due to the investigation of the novel species. Candida africana and Candida dubliniensis are new members of C. albicans complex that are currently gaining both clinical and epidemiologic significance. This study reports the prevalence of C. africana among the strains isolated from patients, by using HWP1 gene size polymorphism.

A total of 235 yeasts confirmed as C. albicans complex based on chromogenic media and ITS-sequencing isolated from various clinical forms of invasive and non-invasive candidiasis mainly candidemia, were re-identified based on HWP1 gene polymorphisms. The Hwp1-PCR amplicons were re-confirmed by sequencing and BLAST analysis.

Based on the HWP1 gene size polymorphism, 223 strains were identified as C. albicans (94.89%) from which 7 isolates produced two DNA fragments (850 and 941 bp). C. dubliniensis (n=4, 1.7%), C. africana (n=1, 0.42%) and mix of C. albicans and C. africana (n=7, 2.97%) were also identified.

Although C. albicans remains the most common Candida species, C. dubliniensis and C. africana are rarely found among the patient isolates. Due to limited information on the molecular epidemiology of this novel yeast, more studies using molecular methods are recommended.

The most common etiological agents of human dermatophytosis in various parts of the world are Trichophyton rubrum, Trichophyton interdigitale, and Epidermophyton floccosum. The main aim of this study was to design and evaluate a simple and straightforward multiplex polymerase chain reaction (PCR)assay for reliable identification/differentiation of these species in clinical isolates.

The reliable sequences of several molecular targets of dermatophytes species were used to design a multiplex PCR for the identification of common pathogenic dermatophytes. The isolates and clinical specimens examined in this study included seven standard strains of dermatophytes, 101 isolates of dermatophytes and non-dermatophyte molds/yeasts which had already been identified by sequencing or PCR-restriction fragment length polymorphism (RFLP), and 155 clinical samples from patients suspected of cutaneous mycoses.

Species-specific primer pairs for T. rubrum and T. interdigitale/T. mentagrophytes were designed based on the sequence data of the translation elongation factor 1-alpha gene, and the primers for E. floccosum targeted the specific sequence of the internal transcribed spacer region (ITS). The multiplex PCR successfully detected T.rubrum, T. interdigitale/T. mentagrophytes, and E. floccosum strains that were identified by sequencing or PCR-RFLP. However, the primer pairs selected for T. interdigitale/T. mentagrophytes cross-reacted with Trichophyton tonsurans. In testing the PCR system directly for clinical samples, the proportion of positive multiplex PCR was higher than positive culture (68.1% vs. 55.4%, respectively).

The multiplex assay could detect three common agents out of several causal agents of dermatophytosis, namely T. rubrum, T. interdigitale, and E. floccosum.Therefore, by adding pan-dermatophyte primers it can be used as a comprehensive detection/identification test.

Detection of coronavirus disease 2019 (COVID-19) in early stage is indispensible for outcome improvement and interruption of transmission chain. Clear understanding of the nature of the diagnostic tests for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and their challenges, collecting the most diagnostically valuable specimen at the right time from the right anatomic site, and interpretation of their findings is important. This review scrutinizes current challenges and interpretation of reverse transcriptase polymerase chain reaction (RT-PCR), as the reference method, loop-mediated isothermal amplification (LAMP), antibody and antigen detection, typical lung imaging characteristics and prominent abnormal changes in laboratory findings of patients with proven COVID-19, and describes how the results may vary over time. Bronchoalveolar lavage fluid and sputum specimens demonstrate the highest positive rates (93% and 72 %, respectively) in molecular diagnosis of COVID-19. Alternatively, repeated RT-PCR assays can be performed; as over time, it is an increase in the likelihood of the SARS-CoV-2 being present in the nasopharynx. Combining clinical evidence with results of chest computed tomography (CT) and RT-PCR can minimize the risk of diagnostic errors. Elevated levels of interleukin 6 (IL‐6) and D-dimer are thought to be closely associated with the occurrence of severe COVID‐19 in adults, and their combined detection can serve as early factors predicting the severity of COVID‐19. Moreover, elevated acute phase proteins are associated with a poor outcome in COVID-19. Serological diagnosis also is an important tool to understand the extent of COVID-19 in the community, and to identify individuals, who are immune. Antibodies begin to increase from the second week of symptom onset.

Leishmaniasis is an expanding neglected tropical disease in the world reporting from 98 countries including Iran. This study focused on eco-epidemiological determinants of the disease following a rapid and unexpected increase of leishmaniasis incidence in a strategic residential district in North-East of Isfahan County, Iran.

This study was accomplished from Apr 2012 to Jan 2014 in a strategic residential zone in North-East of Isfahan County, Esfahan, Iran. Monthly activity, parity, Leishmania infection and susceptibility tests, were determined on sand flies. Some portion of inhabitants and school children were surveyed to find active or passive cases of leishmaniasis and also wild rodents were collected to determine reservoir host.

Totally 5223 sand flies belonging to Phlebotomus and Sergentomyia genus were collected and identified; Ph. papatasi was the dominant species and started to appear in May and disappeared in Oct. The majority of living dissected sand flies were unfed and parous. Ph. papatasi showed 4.6% Leishmania infection through direct examination and 39.54% by nested-PCR respectively. Phlebotomus papatasi was susceptible against deltametrin 0.05%. Totally 2149 people were surveyed and incidence and prevalence of zoonotic cutaneous leishmaniasis estimated as 45.39 and 314.40 per 1000 population. Rodents showed 73.91% and 80% Leishmania infection by direct examination and nested-PCR respectively.

Cutaneous leishmaniasis due to L. major has been established in this area. Rodent control operation and personal protection are highly recommended to control the disease in this focus.

Emergence of Severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) infection (coronavirus disease-2019 or COVID-19) in China since late 2019 and its global distribution has attracted a big international attention. As 2019-nCov is an emerging pathogen, extending our knowledge about the similarities/differences of this virus with other members of the Coronaviridae family, its genetic characteristics, and pathogenesis could be beneficial to develop better strategies of diagnosis, control, and treatment. This pandemic disease is still so new to be fully identified, and there is a long way ahead for complete characterization of SARS-COV-2 and its disease.

 In this study, we investigated the most recent publications from December 2019 to April 2020 on general and genetic features of SARS-COV-2, evaluating the results with primarily approved data about coronaviruses.

 Genetic analysis of SARS-CoV-2 comparing with primarily introduced species indicated high similarity with SARS and Middle East Respiratory Syndrome (MERS) indicating bat source of the emergence.

 Approving the molecular biologic and pathologic similarities between SARS-CoV-2 and previously studied viruses provides prospective for pathogenesis, therapeutics, and development of vaccines, which is more valuable in emergencies of this new pandemic infection.

Invasive aspergillosis (IA) of the central nervous system (CNS) is a devastating complication which is rarely reported in immunocompromised children. In this case presentation, we reported a rare and fatal IA with spinal cord involvement in a 10-year-old child with X-linked chronic granulomatosis disease (CGD).

The child had a previous history of pulmonary tuberculosis. A cervical spine X-ray revealed the involvement of cervical vertebrae (T4/T5) and ribs causing spinal cord compression and epidural abscess. The patient underwent a decompressive laminectomy and mass removal. The histopathology and culture results suggested IA. Despite the aggressive and prolonged therapy, he died within one year. Aspergillus nidulans was identified as the causative agent based on morphological and molecular studies.

This synopsis represents the aggressive behavior of infection caused by A. nidulans in the CGD patient.

Culture-based identification methods have been the gold standard for the diagnosis of candidal onychomycosis. Molecular technologies, such as polymerase chain reaction (PCR) assays, can provide an alternative for the rapid detection of Candida species. The present study was conducted to investigate a pan-Candida PCR assay based on the translation elongation factor 1-alpha (TEF-1α) gene for the detection of the most prevalent pathogenic Candida species.

For the purpose of the study, an optimized pan-Candida PCR primer pair was designed, and the target was amplified and sequenced. The analytical and clinical diagnostic performance of the designed primers was tested using 17 reference strains, 137 nail scrapings suspected of onychomycosis, and 10 healthy nail specimens.

The use of the universal pan-Candida primers designed on TEF-1α gene resulted in the successful amplification of a 270-base pair fragment in all Candida species tested, except for C. glabrata, and reacted neither with other fungi nor with E. coli. The sequence difference count matrix showed poor insertion/deletion differences (0-2 nt) among Candida species. Among 137 nail specimens, 35% (n=48), 30.7% (n=42), and 40.1% (n=55) of the samples were found to be positive by direct microscopy, culture, and pan-Candida PCR, respectively.

Based on the findings, the PCR-based detection targeting the DNA TEF-1α gene is a rapid and simple procedure for the diagnosis of candidal onychomycosis directly from nail sample.

Visceral and cutaneous leishmaniasis are common in some areas of Iran and consider as health problems. Phlebotomus alexandri has been incriminated as a suspected vector for the both form of leishmaniasis.

This study was carried out in 4 western provinces of Iran. Sand flies were collected using sticky traps and light traps from indoor and outdoor resting places. Nested PCR was employed to detect Leishmania parasites among collected sand flies.

Seven hundred and twenty two P. alexandri females were collected and pooled in 179 batches. Results of nested PCR showed, out of 9 samples from East Azerbaijan Province, only one sample was infected by Leishmania infantum. Of 34 individual and pooled samples from Kermanshah Province, only one pooled sample was infected with Leishmania major and among 30 individual and pooled samples in Fars Province, five specimens were infected by L. major, L. infantum, Leishmania donovani and Leishmania tropica. Furthermore, out of 108 individual and pooled samples from Khuzestan Province, 10 samples showed infection with L. major and L. infantum.

The results of this study showed that P. alexandri is more active in hot zones than in moderate zones and this species may be considered as a permissive species.

Dermatophytes create the most common fungal disease in humans, called dermatophytosis. The two species of Trichophyton rubrum and Trichophyton interdigital are responsible for over 80% of types of dermatophytosis. So far, several morphological and physiological methods have been used to differentiate these very similar species, but these methods are generally time-consuming and have low specificity. The purpose of this study was to introduce a simple and rapid duplex polymerase chain reaction (PCR) reaction to differentiate these two species from each other. This research was an analytical and experimental study that was carried out from 2017 to 2018 in the Medical Mycology Laboratory, School of Public Health, Tehran University of Medical Sciences, Iran. For this purpose, the nucleotide sequences of the 4 regions of internal transcribed spacer (ITS), beta-tubulin, elongation factor 1 alpha and calmodulin in the two considered species of fungi were conducted bioinformatics analysis. The differences and similarities of nucleotides between two species in each of these genes were studied for selecting the primer. The specificity of selected primers was tested for duplex PCR reaction against sequenced isolates of dermatophyte species. According to the total data, the specific primers were selected from elongation factor 1 alpha gene. These primers produced a product of 173 and 384 bp, in Trichophyton rubrum and Trichophyton interdigital, respectively. They had high specificity in the face of various dermatophytes. The length of nucleotide sequences found in the genebank of this gene in the two species is between 700 and 770 bp. The similarity of the two species in this region is 94.6% and differs by 78 bp. Of the 107 extracted DNAs from clinical dermatophyte isolates, in duplex PCR 24 isolates were positive with Trichophyton interdigital primer and 71 isolates against Trichophyton rubrum. The remaining isolates, which included 6, were negative in this reaction, which included other dermatophyte species. This method is a specific and fast differential method compared to conventional methods for identifying Trichophyton rubrum and Trichophyton interdigital from each other.

Mucormycosis is an acute and invasive fungal infection with a high mortality rate. Mucorales are less sensitive than other types of fungi to most antifungal agents. Amphotericin B (AMB) is one treatment option for this infection, but in recent studies, the antifungal activity of statins against Mucorales was shown. Therefore, therapy that combines AMB with these agents may have better effects in management of patients with mucormycosis. We evaluated the in vitro activity of AMB alone and in combination with statins, against Mucorales. Susceptibility profiles of AMB alone and in combination with two statins, atorvastatin (ATO) and lovastatin (LOV) determined against clinical (n: 15) and environmental (n: 5) Rhizopus oryzae isolates, obtained between Jan 2009 and Oct 2016 from patients with uncontrolled diabetes mellitus and cancer referred to the Department of Parasitology and Medical Mycology of Tehran University of Medical Sciences, Tehran, Iran. It was performed by microdilution method, based on the Clinical and Laboratory Standard Institute (CLSI) M38-A2 guideline. All clinical and environmental isolates were susceptible to AMB (MIC≤1 µg/mL). The results of the interactions between AMB and the two statins were positive. The AMB-ATO (GM: 0.13 µg/Ml) combination produced greater activity than the AMB-LOV (GM: 0.26 µg/mL) combination. AMB, in combination with ATO and LOV, reacts positively against clinical and environmental R. oryzae isolates. This combination strategy may lead to more effective treatment of mucormycosis and fewer side effects using low dose of AMB.

Accurate speciation of the clinical yeast isolates is essential to a targeted treatment. The aim of this study was to identify and determine the frequency of uncommon and rare yeast species causing fungal infections that may be misidentified. During a 10-month period, yeast isolates collected from patients referring to or hospitalized in some educational hospitals in Tehran, Iran, were included in this study. In addition to conventional methods such as direct microscopy and culture on CHROMagar Candida, molecular methods including PCR-RFLP and ITS-sequencing were used for the accurate identification of the yeast strains. Among 930 yeast isolates recovered from normally sterile and other clinical specimens, a total number of 27 strains were identified as uncommon Candida species and three were identified as rare non-Candida species yeasts. They included C. kefyr (n = 12), C. lusitaniae (n = 8), C. intermedia (n = 3), C. orthopsilosis, C. guilliermondii, and Trichosporon asahii (each n = 2), and Magnusiomyces capitatus (n = 1). The isolation of less common or rare yeast species, which can cause a variety of infections from superficial to systemic infections, is increasingly reported. Since this uncommon yeast species may exhibit low susceptibility to some antifungal agents, the use of reliable methods for accurate screening of these yeasts is necessary.

Mucormycosis is a life-threatening infection due to the members of the Mucorales order with increasing incidence during the last decades. Rhizopus spp. are the most common causes of disease; however, this infection could be developed by various other species. This study presented the clinical features and predisposing factors of several patients with mucormycosis along with the causative agents using sequence analysis. Clinical specimens of 25 cases with mucormycosis were included in this study. Direct examination and culture were performed for all specimens and then isolated fungi were identified based on their morphology and sequence analysis of ribosomal DNA. The patients were comprised of 17 (68%) females and 8 (32%) males with the mean age of 47.16 ± 17.4 years. Rhino-cerebral mucormycosis was the most common clinical form (24 cases) followed by pulmonary mucormycosis (one case). Diabetes mellitus was the most common predisposing factor (n = 17, 68%). The culture was positive in 15 specimens and the isolated fungi were morphologically identified as Rhizopus spp., subsequently, all of the isolates were identified as Rhizopus oryzae at the species level using the molecular method. Diabetes and R. oryzae played the most important roles as the predisposing factor and etiologic agent of mucormycosis, respectively.

Candiduria in hospitalized patients may represent contamination, colonization, or urinary tract in-fections. On the other hand, candidemia and upper urinary tract infection could be the complications of can-diduria. The aim of this study was to determine candiduria in hospitalized patients and identify isolated Candida species by conventional and molecular methods. This cross-sectional study was conducted on hospitalized patients in Imam Khomeini and Mostafa Khomeini hospitals in Ilam, western Iran from Jan to Dec 2016. Urine samples of hospitalized patients were col-lected during a period of 4 months for diagnosis of candiduria. Primary identification was done by conventional methods. PCR profile was carried out using phenol-chloroform method and confirmed using restriction fragment length polymorphism (PCR-RFLP) technique by MspI restriction enzyme. Candiduria was diagnosed in 18 (9.2%) cases from a total of 195 patients. Isolated yeasts were identified as C. albicans (n: 13), C. glabrata (n: 5), and C. parapsilosis (n: 1) in the one case both C. albicans and C. glabrata were isolated from a urine sample. Candida urinary tract infection is becoming increasingly common in hospitalized patients but, differentia-tion fungal colonization from infection and identification of etiologic agents for optimal treatment is necessary.

Toxoplasma gondii, cause severe medical complications in infants and immune-compromised individuals. As using early, sensitive and rapid technique has major in diagnosis of toxoplasmosis, the present study was aimed to detect parasite by using from repetitive element (RE) and B1genes, in blood samples of seropositive immunocompromised patients and pregnant women. A total of 110 peripheral blood samples were collected from seropositive cases with anti-T. gondii antibodies, including immunocompromised patients and pregnant women. DNA was extracted by a commercial kit and subjected to TaqMan probe-based real-time PCR assay by using primers and probes specific for RE and B1 genes, separately. The data were analyzed by Kappa test and SPSS-22 software. In the pregnant women, 17 (68%) and 14 (56%) samples from 25 IgM+/ IgG+ cases and, 7 (25%) and 6 (21.4%) samples from 28 IgG+/IgM- cases were positive by RE and B1 real time PCR, respectively. Likewise, in immunocompromised group, 20 (66.6%) and 17 (56.6%) samples from 30 IgM+/ IgG+ cases and 2 (7.4%) and 2 (7.4%) samples from 27 IgG+/ IgM- cases were positive by RE and B1 real time PCR, respectively. Probe-based real time PCR assay is a quantitative approach for early diagnosis of T. gondii infection in clinical samples. Moreover, this method can be more appropriate in diagnosis of acute and reactivated toxoplasmosis. In addition our results indicated that RE gene is more sensitive than B1 gene.

The aspects of the epidemiology of bloodstream Candida infections including clinical features, the causal agents, underlying conditions, and risk factors have not been well-defined in Iranian pediatric patients. The aim of this observational study was to identify uncommon Candida species isolated from blood and other normally sterile specimens of the neonates and children admitted to intensive care units at Children’s medical center, Tehran, Iran.
The study was carried out prospectively on patients A total of 16 out of 136 isolates were recognized as uncommon or rare Candida species. According to the sequence analysis, these isolates were identified as C. orthopsilosis (N = 5, 3.7%), C. glabrate (n = 4, 2.5%), C. dubliniensis (n = 2, 1.5%), C. lusitaniae (n = 2, 1.5%), C. kefyr (n = 2, 1.5%), and C. intermedia (n = 1, 0.75%)
Candida species, which were once considered harmless, have now been recognized as causative agents of candidemia. It is essential to consider, manage, and control the conditions that lead to the development of these unusual but severe cases of candidemia.

Background and Candida parapsilosis is a common cause of candidemia in children and onco-hematologic patients as well as in patients with septic arthritis, peritonitis, vaginitis, and nail and skin infections. Here, we evaluated intra- and inter-species beta tubulin DNA sequence variation within the C. parapsilosis complex with a view to establishing its utility in the identification and phylogenetic analysis of the species.
The novel primers successfully amplified the target for all 23 strains included in the study.
Results and Multiple alignment of 623–629 bp-long nucleotide (nt) sequences reflecting the beta tubulin gene indicated an inter-species divergence ranging from 0 to 68 nt in C. parapsilosis, C. orthopsilosis and C. metapsilosis with a mean similarity of 84.7% among the species. Meanwhile, intra-species differences of 0–20 and 0–6 nt were found within the strains of C. parapsilosis and C. orthopsilosis, respectively. The phylogenetic tree topology was characterized by a clade made up by C. parapsilosis and C. orthopsilosis, while C. metapsilosis formed a related but separate lineage. Our data provide the basis for further discoveries of the relationship between the species belonging to the C. parapsilosis complex and the value of beta tubulin DNA sequence data in the identification and taxonomy of C. parapsilosis and other pathogenic yeasts.

Candida species originating from either endogenous or exogenous sources are one of the main causes of opportunistic infections. Colonization is an important independent risk factor for invasive candidiasis, and many patients admitted to neonatal intensive care unit (NICU) and pediatric intensive care unit (PICU) are colonized with Candida species that may result in invasive candidiasis. Awareness among clinicians about various aspects of colonization is critical to optimal management. The aim of this study was to determine the frequency and species distribution of Candida strains isolated from predisposed patients hospitalized at Children’s Medical Center (CMC), Tehran, Iran.
From June 2014 to June 2016, 347 Candida isolates were collected from 341 patients either hospitalized in different wards or referred as outpatients. The yeasts were identified by colony color characteristics using CHROMagar Candida medium and by amplification of the ITS1-5.8S rDNA-ITS2 region in DNA extracted from each isolate followed by analysis of species specific electrophoretic patterns of PCR products digested with the restriction enzyme MspI.
Of the 341 patients, 213 were males and 128 were females. Most samples were obtained from the 1 – 12-month age group, and the majority of samples represented urine (n = 182), throat swabs (n = 57), and stool samples (n = 53), respectively. The samples were mostly from patients in general wards. The most commonly isolated species was C. albicans (77%), followed by C. tropicalis (8.4%), C. parapsilosis (7.5%), C. glabrata (2.3%), C. kefyr (1.7%), C. krusei (1.1%), C. lusitaniae (0.6%), C. guilliermondii (0.3%), C. albicans C. parapsilosis (1.4%), and C. albicans C. glabrata (0.3%).
C. albicans is the most common species isolated from children in Iran, followed by C. tropicalis and C. parapsilosis, a prevalence pattern that is relatively different from studies in other countries. Neonates and infants 1 - 12 months of age hospitalized in ICU, were more colonized by Candida species than other groups.