ABSTRACT
Background and Aim: The emergence of nonfermenter bacteria that are resistant to multidrug resistant ESBL are nowadays a principal problem for hospitalized patients. The present study aimed at surveying the emergence of nonfermenter bacteria resistant to multi-drug ESBL producing isolated from patients blood samples using BACTEC 9240 automatic system in Shiraz
Materials and Methods: In this cross-sectional study, 4825 blood specimens were collected from hospitalized patients in Shiraz [Iran], and positive samples were detected by means of BACTEC 9240 automatic system. The isolates containing nonfermenter bacteria were identified based on biochemical tests embedded in the API-20E system. Antibiotic sensitivity test was performed and identification of ESBL producing strains were done using phenotypic detection of extended spectrum beta-lactamase producing isolates [DDST] according to CLSI [2013]guidelines
Results: Out of 4825 blood samples, 1145 [24%] specimen were gram-positive using BACTEC system
Among all isolated microorganisms, 206 isolates were non-fermenting gram- negative bacteria
The most common non-fermenter isolates were Pseudomonas spp. [48%], Acinetobacter spp. [41.7%] ,and Stenotrophomonas spp. [8.2%]. Seventy of them [81.4%] were Acinetobacter spp. which were ESBL positive. Among beta-lactam antibiotics, Pseudomonas spp. showed the best sensitivity to piperacillintazobactam [46.5%]
Conclusion: It was found that ?-lactam antibiotics are not effective against more than 40% of Pseudomonas spp. infections and 78% Acinetobacter infections. Emergence of multi-drug resistant strains that are resistant to most antibiotic classes is a major public health problem in Iran. To resolve this problem using of practical guidelines is critical
ABSTRACT
Accurate and rapid diagnosis of human cytomegalovirus [HCMV] disease in immunocompromised patients has remained as a challenge. Quantitative competitive PCR [QC-PCR] methods for detection of HCMV in these individuals have improved the positive and negative predictive values of PCR for diagnosis of HCMV disease. In this study we used QC-PCR assay, using a co-amplified DNA standard, to quantitate the HCMV glycoprotein B [gB] gene in different samples. A DNA internal standard [IS] was designed by replacing HCMV primer binding site at 5' ends of primers that amplifies a 156-bp fragment of lambda genome, and a 200 bp amplicon was produced. Two DNA fragments of 257 bp wild type and 200-bp [IS] were co-amplified with the same oligonucleotide primer sets, analyzed by gel electrophoresis and used for construction of a standard curve. From this, the copy number of the gB gene present in different samples could be determined. Co-amplification with 1,000 copies of IS, allowed quantitation of 10-100,000 of HCMV DNA in a single PCR. This rapid assay avoids using radioactive components and other less efficient quantitative systems. It has the potential for early identification of patients at high risk of development of HCMV disease, and is useful for therapeutic monitoring