scholarly journals Reliable and Sensitive Nested PCR for the Detection of Chlamydia in Sputum

2021 ◽  
Vol 9 (5) ◽  
pp. 935
Author(s):  
Martina Smolejová ◽  
Iveta Cihová ◽  
Pavol Sulo
Keyword(s):  
Nested Pcr ◽  
Chlamydia Pneumoniae ◽  
Dna Degradation ◽  
Atypical Pneumonia ◽  
Intracellular Pathogens ◽  
Human Pathogen ◽  
Three Samples ◽  
Pcr Products ◽  
Low Sensitivity ◽  
Sick Individual

Chlamydia are Gram-negative, intracellular pathogens colonizing epithelial mucosa. They cause primarily atypical pneumonia and have recently been associated with chronic diseases. Diagnostics relies almost exclusively on serological methods; PCR tests are used rarely because in patients with positive ELISA, it is nearly impossible to identify chlamydial DNA. This paradox is associated with DNA degradation in sputum samples, low abundance, and low sensitivity of PCR systems. In a newly designed and validated “nested” PCR (NPCR) assay, it was possible to amplify DNA of Chlamydia known to infect humans in 31% samples. The reliability of the assay was confirmed by DNA sequencing, and all PCR products belonged exclusively to the Chlamydiales, mainly recognized as Chlamydia pneumoniae. Three samples were related to Ca. Rhabdochlamydia porcellionis and Ca. Renichlamydia lutjani, which infect arthropods. In one case, samples were taken from sick individual, indicating the potential as a human pathogen.

scholarly journals First Report of a Group 16SrII Phytoplasma Infecting Chickpea in Oman

Plant Disease ◽  
2006 ◽  
Vol 90 (7) ◽  
pp. 973-973 ◽  
Author(s):  
N. A. Al-Saady ◽  
A. M. Al-Subhi ◽  
A. Al-Nabhani ◽  
A. J. Khan
Keyword(s):  
Nested Pcr ◽  
Animal Feed ◽  
Restriction Enzymes ◽  
Universal Primers ◽  
Polymorphism Analysis ◽  
Disease Symptoms ◽  
First Report ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Group 2

Chickpea (Cicer arietinum), locally known as “Dungo”, is grown for legume and animal feed mainly in the interior region of Oman. During February 2006, survey samples of chickpea leaves from plants showing yellows disease symptoms that included phyllody and little leaf were collected from the Nizwa Region (175 km south of Muscat). Total nucleic acid was extracted from asymptomatic and symptomatic chickpea leaves using a cetyltrimethylammoniumbromide method with modifications (3). All leaf samples from eight symptomatic plants consistently tested positive using a polymerase chain reaction assay (PCR) with phytoplasma universal primers (P1/P7) that amplify a 1.8-kb phytoplasma rDNA product and followed by nested PCR with R16F2n/R16R2 primers yielding a product of 1.2 kb (2). No PCR products were evident when DNA extracted from healthy plants was used as template. Restriction fragment length polymorphism analysis of nested PCR products by separate digestion with Tru9I, HaeIII, HpaII, AluI, TaqI, HhaI, and RsaI restriction enzymes revealed that a phytoplasma belonging to group 16SrII peanut witches'-broom group (2) was associated with chickpea phyllody and little leaf disease in Oman. Restriction profiles of chickpea phytoplasma were identical with those of alfalfa witches'-broom phytoplasma, a known subgroup 16SrII-B strain (3). To our knowledge, this is the first report of phytoplasma infecting chickpea crops in Oman. References: (1) A. J. Khan et al. Phytopathology, 92:1038, 2002. (2). I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998 (3) M. A. Saghai-Maroof et al. Proc. Natl. Acad. Sci. USA. 81:8014, 1984.

scholarly journals First Report of a Group 16SrII Phytoplasma Associated with Witches'-Broom of Eggplant in Oman

Plant Disease ◽  
2011 ◽  
Vol 95 (3) ◽  
pp. 360-360 ◽  
Author(s):  
A. M. Al-Subhi ◽  
N. A. Al-Saady ◽  
A. J. Khan ◽  
M. L. Deadman
Keyword(s):  
Nested Pcr ◽  
Solanum Melongena ◽  
Positive Control ◽  
Rrna Gene ◽  
Direct Pcr ◽  
First Report ◽  
Pcr Product ◽  
Pcr Products ◽  
Broom Disease ◽  
Phytoplasma Infection

Eggplant (Solanum melongena L.) belongs to the family Solanaceae and is an important vegetable cash crop grown in most parts of Oman. In February 2010, plants showing phyllody symptoms and proliferation of shoots resembling those caused by phytoplasma infection were observed at Khasab, 500 km north of Muscat. Total genomic DNA was extracted from healthy and two symptomatic plants with a modified (CTAB) buffer method (2) and analyzed by direct and nested PCR with universal phytoplasma 16S rDNA primers P1/P7 and R16F2n/ R16R2, respectively. PCR amplifications from all infected plants yielded an expected product of 1.8 kb with P1/P7 primers and a 1.2-kb fragment with nested PCR, while no products were evident with DNA from healthy plants. Restriction fragment length polymorphism (RFLP) profiles of the 1.2-kb nested PCR products of two eggplant phyllody phytoplasma and five phytoplasma control strains belonging to different groups used as positive control were generated with the restriction endonucleases RsaI, AluI, Tru9I, T-HB8I, and HpaII. The eggplant phytoplasma DNA yielded patterns similar to alfalfa witches'-broom phytoplasma (GenBank Accession No. AF438413) belonging to subgroup 16SrII-D, which has been recorded in Oman (1). The DNA sequence of the 1.8-kb direct PCR product was deposited in GenBank (Accession No. HQ423156). Sequence homology results using BLAST revealed that the eggplant phyllody phytoplasma shared >99% sequence identity with Scaevola witches'-broom phytoplasma (Accession No. AB257291.1), eggplant phyllody phytoplasma (Accession No. FN257482.1), and alfalfa witches'-broom phytoplasma (Accession No. AY169323). The RFLP and BLAST results of 16S rRNA gene sequences confirm that eggplant phyllody phytoplasma is similar to the alfalfa phytoplasma belonging to subgroup 16SrII-D. To our knowledge, this is the first report of a phytoplasma of the 16SrII-D group causing witches'-broom disease on eggplant in Oman. References: (1) A. J. Khan et al. Phytopathology 92:1038, 2002. (2) M. A. Saghai-Maroof et al. Proc. Natl. Acad. Sci. USA, 81:8014, 1984.

scholarly journals First Report of an Aster Yellows Phytoplasma Associated with Cabbage in Southern Texas

Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 447-447 ◽  
Author(s):  
I.-M. Lee ◽  
R. A. Dane ◽  
M. C. Black ◽  
Noel Troxclair
Keyword(s):  
16S Rdna ◽  
Nested Pcr ◽  
Restriction Enzymes ◽  
Diagnostic Procedures ◽  
Acid Extraction ◽  
Insect Vector ◽  
Aster Yellows ◽  
First Report ◽  
Pcr Products ◽  
Aster Yellows Phytoplasma

In early spring 2000 carrot crops in southwestern Texas were severely infected by an outbreak of phyllody associated with aster yellows phytoplasma. Cabbage crops that had been planted adjacent to these carrot fields began to display previously unobserved symptoms characteristic of phytoplasma infection. Symptoms included purple discoloration in leaf veins and at the outer edges of leaves on cabbage heads. Proliferation of sprouts also occurred at the base of the stem and between leaf layers of some plants, and sprouts sometimes continued to proliferate on extended stems. About 5% of cabbage plants in the field exhibited these symptoms. Two symptomless and four symptomatic cabbage heads were collected in early April from one cabbage field. Veinal tissues were stripped from each sample and used for total nucleic acid extraction. To obtain specific and sufficient amount of PCR products for analysis, nested PCR was performed by using primer pairs (first with P1/P7 followed by R16F2n/R16R2) (1,2) universal for phytoplasma detection. A specific 16S rDNA fragment (about 1.2 kb) was strongly amplified from the four symptomatic but not from the two asymptomatic samples. The nested PCR products obtained from the four symptomatic samples were then analyzed by restriction fragment length polymorphism (RFLP) using the restriction enzymes MseI, HhaI, and HpaII, and the RFLP patterns were compared to the published patterns of known phytoplasmas (1). The resulting RFLP patterns were identical to those of a phytoplasma belonging to subgroup B of the aster yellows phytoplasma group (16SrI). These RFLP patterns were also evident in putative restriction sites observed in a 1.5 kbp nucleotide sequence of the 16S rDNA. This is the first report of aster yellows phytoplasma associated disease symptoms in cabbage in Texas. The occurrence of cabbage proliferation coincided with the presence of high populations of the insect vector, aster leafhopper. References: (1) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) B. Schneider et al. 1995. Molecular and Diagnostic Procedures in Mycoplasmology, Vol. I. Academic Press, San Diego, CA.

scholarly journals Circulation of bovine viral diarrhea virus – 1 (BVDV-1) in dairy cattle and buffalo farms in Ismailia Province, Egypt

2015 ◽  
Vol 9 (12) ◽  
pp. 1331-1337 ◽  
Author(s):  
Mohamed Ahmed Soltan ◽  
Rebecca P Wilkes ◽  
Mohamed Nagy Elsheery ◽  
Mahmoud Mohy Elhaig ◽  
Matthhew C Riley ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Dairy Cattle ◽  
Real Time ◽  
Bovine Viral Diarrhea ◽  
Rt Pcr ◽  
Cattle Farm ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Three Samples ◽  
Pcr Products

Introduction: Bovine viral diarrhea (BVD) is one of the most economically significant diseases in the bovine industry causing losses due to diarrhea, reproductive disorders, immunosuppression and mortalities. The aim of our investigation was to detect and subtype BVDV from calves on two dairy cattle and two buffalo farms in Ismailia province, Egypt as an indicator of BVDV infection status in the province. Methodology: A total of 298 blood samples were collected and tested using an optimized one-step, real-time multiplex Taqman-based RT-PCR. All the positive samples by the multiplex real-time RT-PCR were tested using conventional RT-PCR to amplify multiple areas of the genome for further phylogenetic analysis and subtyping. Results: Thirty one (10.4%) of the tested samples were positive for BVDV-1. Only three samples, all from a single dairy cattle farm, had enough viral RNA to be amplified by RT-PCR. The PCR products were sequenced and phylogenetic analysis revealed detection of BVDV-1b. The detected strain is closely related to worldwide BVDV-1b strains, making it difficult to trace its origin. Nucleotide and amino acid alignments of the E2 glycoprotein region of the detected strain with other BVDV-1b strains showed high divergence, with identity ranging from 81.3% to 93.6% and 85.3% to 93.6%, respectively. Conclusion: To our knowledge, this is the first report describing the circulation of BVDV-1b in Egyptian dairy cattle populations.

scholarly journals Detection and Discrimination of Cryptosporidium parvum and C. hominis in Water Samples by Immunomagnetic Separation-PCR

2005 ◽  
Vol 71 (2) ◽  
pp. 898-903 ◽  
Author(s):  
Yoshitsugu Ochiai ◽  
Chieko Takada ◽  
Mitsugu Hosaka
Keyword(s):  
Cryptosporidium Parvum ◽  
Water Samples ◽  
Nested Pcr ◽  
Fragment Length Polymorphism ◽  
Detection Method ◽  
Fluorescent Antibody ◽  
Immunomagnetic Separation ◽  
Antibody Assay ◽  
Pcr Products ◽  
Genetic Detection

ABSTRACT Cryptosporidium parvum and C. hominis have been the cause of large and serious outbreaks of waterborne cryptosporidiosis. A specific and sensitive recovery-detection method is required for control of this pathogen in drinking water. In the present study, nested PCR-restriction fragment length polymorphism (RFLP), which targets the divergent Cpgp40/15 gene, was developed. This nested PCR detected only the gene derived from C. parvum and C. hominis strains, and RFLP was able to discriminate between the PCR products from C. parvum and C. hominis. To evaluate the sensitivity of nested PCR, C. parvum oocysts inoculated in water samples of two different turbidities were recovered by immunomagnetic separation (IMS) and detected by nested PCR and fluorescent antibody assay (FA). Genetic detection by nested PCR and oocyst number confirmed by FA were compared, and the results suggested that detection by nested PCR depends on the confirmed oocyst number and that nested PCR in combination with IMS has the ability to detect a single oocyst in a water sample. We applied an agitation procedure with river water solids to which oocysts were added to evaluate the recovery and detection by the procedure in environmental samples and found some decrease in the rate of detection by IMS.

scholarly journals Assaying Chlamydia pneumoniae Persistence in Monocyte-Derived Macrophages Identifies Dibenzocyclooctadiene Lignans as Phenotypic Switchers

Molecules ◽  
2020 ◽  
Vol 25 (2) ◽  
pp. 294 ◽  
Author(s):  
Eveliina Taavitsainen ◽  
Maarit Kortesoja ◽  
Tanja Bruun ◽  
Niklas G. Johansson ◽  
Leena Hanski
Keyword(s):  
Chlamydia Pneumoniae ◽  
Concomitant Administration ◽  
Productive Infection ◽  
Intracellular Bacteria ◽  
Bacterial Eradication ◽  
Human Pathogen ◽  
Quantitative Culture ◽  
Phenotypic Switch ◽  
Host Interactions ◽  
Adjuvant Therapies

Antibiotic-tolerant persister bacteria involve frequent treatment failures, relapsing infections and the need for extended antibiotic treatment. The virulence of an intracellular human pathogen C. pneumoniae is tightly linked to its propensity for persistence and means for its chemosensitization are urgently needed. In the current work, persistence of C. pneumoniae clinical isolate CV6 was studied in THP-1 macrophages using quantitative PCR and quantitative culture. A dibenzocyclooctadiene lignan schisandrin reverted C. pneumoniae persistence and promoted productive infection. The concomitant administration of schisandrin and azithromycin resulted in significantly improved bacterial eradication compared to sole azithromycin treatment. In addition, the closely related lignan schisandrin C was superior to azithromycin in eradicating the C. pneumoniae infection from the macrophages. The observed chemosensitization of C. pneumoniae was associated with the suppression of cellular glutathione pools by the lignans, implying to a previously unknown aspect of chlamydia–host interactions. These data indicate that schisandrin lignans induce a phenotypic switch in C. pneumoniae, promoting the productive and antibiotic-susceptible phenotype instead of persistence. By this means, these medicinal plant -derived compounds show potential as adjuvant therapies for intracellular bacteria resuscitation.

scholarly journals Isolation of Chlamydia pneumoniae Clonal Variants by a Focus-Forming Assay

2002 ◽  
Vol 70 (10) ◽  
pp. 5827-5834 ◽  
Author(s):  
Jens Gieffers ◽  
Robert J. Belland ◽  
William Whitmire ◽  
Scot Ouellette ◽  
Deborah Crane ◽  
...  
Keyword(s):  
Cardiovascular Disease ◽  
Respiratory Infection ◽  
Chlamydia Pneumoniae ◽  
Genetic Differences ◽  
Genomic Sequencing ◽  
Human Pathogen ◽  
Obligate Intracellular ◽  
Respiratory Isolate

ABSTRACT Chlamydia pneumoniae is an obligate intracellular prokaryotic human pathogen that causes community-acquired respiratory infection and has been associated with atherosclerosis and cardiovascular disease. Unexpected results from genomic sequencing indicate that significant intrastrain polymorphism exists for some C. pneumoniae isolates. These polymorphisms could reflect genotypes with differing disease-causing characteristics. A definitive means to test this hypothesis is to obtain genetically homogeneous clonal populations of the pathogen and test them in models of infection and disease. To date, methods for cloning C. pneumoniae have not been reported. In this study, we describe the isolation of clonal variants with genetic differences in the tyrP locus from a polymorphic respiratory isolate, using a novel focus-forming assay. These results now allow investigations on the biology and pathogenesis of C. pneumoniae clonal genovars that could lead to new insights into the pathogenesis of this important human pathogen.

Nested PCR protocol for the rapid detection of Escherichia coli in potable water

1996 ◽  
Vol 42 (8) ◽  
pp. 862-866 ◽  
Author(s):  
David Juck ◽  
Jordan Ingram ◽  
Michèle Prévost ◽  
Josée Coallier ◽  
Charles Greer
Keyword(s):  
Escherichia Coli ◽  
Nested Pcr ◽  
Potable Water ◽  
Test Organism ◽  
Bacterial Cells ◽  
Nested Polymerase Chain Reaction ◽  
Fecal Indicator ◽  
E Coli ◽  
Pcr Products ◽  
Polymerase Chain

A rapid and sensitive method for the detection of low levels of bacteria in potable water was developed. The fecal indicator bacterium Escherichia coli was used as the test organism in a filtration concentration–nested polymerase chain reaction (PCR) protocol, combined with ethidium bromide visualization of PCR products. Two sets of primers were designed from the E. coli specific β-glucuronidase gene (uidA), the primary pair producing a 486-bp fragment that was used as template for the nested primer pair delineating a 186-bp fragment. This protocol can detect 1–10 bacterial cells/50 mL water sample within 6–8 h, in contrast to traditional culturing or Southern hybridization methods which require 2–3 days for results.Key words: nested PCR, sensitive, detection, potable water.

scholarly journals Πνευμονία της κοινότητας

2013 ◽  
Author(s):  
Χαράλαμπος Νεοκλέους
Keyword(s):  
Streptococcus Pneumoniae ◽  
Haemophilus Influenzae ◽  
Legionella Pneumophila ◽  
Nested Pcr ◽  
Chlamydia Pneumoniae ◽  
Haemophilus Influenza ◽  
Streptococcus Pneumonia

Πνευμονία της κοινότητας ορίζεται η πνευμονία που αναπτύσσεται έξω από νοσοκομειακό περιβάλλον, σε άτομο που δεν έχει πρόσφατα νοσηλευθεί και δεν είναι ανοσοκατασταλμένο. Στόχος της παρούσας διατριβής ήταν η μελέτη του είδους και της συχνότητας των παθογόνων βακτηρίων που ενέχονται στην πρόκληση της πνευμονίας της κοινότητας στην περιοχή της Θεσσαλίας, καθώς και η σύγκριση των διαφόρων κλασσικών (Gram χρώση, καλλιέργειες σε κοινά στερεά θρεπτικά μέσα), ορολογικών και νεότερων (μοριακές τεχνικές όπως PCR, nested PCR, ανίχνευση αντιγόνου των Streptococcus pneumoniae και της Legionella pneumophila) μικροβιολογικών μεθόδων που χρησιμοποιούνται για τη διάγνωση αυτών. Συνολικά μελετήθηκαν 215 ενήλικες ασθενείς που νοσηλεύθηκαν με πνευμονία της κοινότητας. Ως διαγνωστικά υλικά χρησιμοποιήθηκαν πτύελα, ορός αίματος, ούρα, και πλευριτικό υγρό. Για την ανίχνευση των συχνότερων παθογόνων μικροοργανισμών που προκαλούν πνευμονία της κοινότητας χρησιμοποιηθήκαν Gram χρώση, καλλιέργειες, PCR και nested-PCR, ορολογικές μέθοδοι και η μέθοδος ανίχνευσης του αντιγόνου του Streptococcus pneumoniae και της Legionella pneumophila στα ούρα. Με τη χρήση όλων των διαγνωστικών μεθόδων ανιχνεύθηκαν 106 μικροοργανισμοί σε 106 (49,3%) ασθενείς. Σε 109 (50,7%) ασθενείς παρόλη τη χρήση όλων των διαγνωστικών δοκιμασιών, ο αιτιολογικός παράγοντας παρέμεινε αταυτοποίητος. Στην συντριπτική πλειοψηφία των ασθενών (37,2%) απομονώθηκαν τυπικά παθογόνα του κατώτερου αναπνευστικού με κυρίαρχα παθογόνα τον Streptococcus pneumonia και τον Haemophilus influenzae. Η συχνότητα απομόνωσης άτυπων βακτηρίων (Μycoplasma pneumoniae, Chlamydia pneumoniae, Legionella pneumophila) στην περιοχή της Θεσσαλίας ήταν ιδιαίτερα αυξημένη (12,1%), γεγονός που δείχνει ότι είναι αναγκαία ως εμπειρική θεραπεία η χρήση αντιβιοτικής αγωγής με σχήματα που περιέχουν μακρολίδες,τετρακυκλίνες ή φθοριοκινολόνες που αποτελούν αντιβιοτικά εκλογής για την αντιμετώπιση των πιο πάνω μικροοργανισμών πέραν των β-λακταμικών που κατά βάση χρησιμοποιούνται.Κυρίαρχο παθογόνο του κατώτερου αναπνευστικού ήταν ο Haemophilus influenzae που ανιχνεύθηκε κυρίως σε ασθενείς με ΧΑΠ οι οποίοι αποτελούσαν σημαντικό ποσοστό του πληθυσμού μελέτης. Από τους 38 Haemophilus influenzae που ανιχνεύθηκαν, με καλλιέργεια πτυέλων και PCR, οι 36 (94,7%) ανιχνεύθηκαν στα πτύελα ασθενών με ΧΑΠ που νοσηλεύθηκαν λόγω πνευμονίας της κοινότητας. Σε 36 (64,3%) ασθενείς με ΧΑΠ ο μικροοργανισμός που ανιχνεύθηκε ήταν ο Haemophilus influenzae. Η συχνότητα των υπολοίπων μικροοργανισμών βρίσκεται σε ευρεία διακύμανση σε σύγκριση με την συχνότητα που αναφέρουν προηγούμενες επιδημιολογικές μελέτες.Ωστόσο, παρά τη χρήση όλων των διαγνωστικών μεθόδων, ο αιτιολογικός παράγοντας παρέμεινε αταυτοποίητος σε περισσότερους από τους μισούς ασθενείς. Ένα μέρος των αταυτοποίητων πνευμονιών πιθανόν να αποτελούν οι ιογενείς πνευμονίες, οι πνευμονίες οφειλόμενες σε μικροοργανισμούς όπως Coxiella burnetti και Chlamydia psittaki οι οποίοι δεν μελετήθηκαν στην παρούσα μελέτη καθώς και ένας αριθμός ασθενών με πνευμονία από άτυπα βακτήρια που η διάγνωση πιθανόν να τίθετο με την ανεύρεση τετραπλασιασμού του τίτλου των IgG αντισωμάτων.Η PCR έναντι της καλλιέργειας πτυέλων πλεονεκτούσε στην ανίχνευση του Haemophilus Influenza στους ασθενείς με ΧΑΠ με πνευμονία της κοινότητας αφού αναδείκνυε παρουσία DNA του μικροοργανισμού στα πτύελα με ταυτόχρονη αρνητική την καλλιέργεια πτυέλων. Οι ορολογικές μέθοδοι υπερτερούσαν της PCR στη διάγνωση των άτυπων παθογόνων του κατώτερου αναπνευστικού. Παρόλα αυτά, η PCR μπορεί να χρησιμοποιηθεί παράλληλα με τις ορολογικές δοκιμασίες ως ταχεία μέθοδος διάγνωσης των άτυπων παθογόνων. Ενώ η μέθοδος ανίχνευσης αντιγόνου του Streptococcus pneumoniae στα ούρα φαίνεται πως δεν προσέφερε σε σημαντικό αριθμό ασθενών επιπρόσθετες πληροφορίες από την PCR, το θετικό αποτέλεσμα αποτελεί ένα επιπλέον στοιχείο που συνηγορεί ότι ο Streptococcus pneumonia που ανιχνεύθηκε με τις υπόλοιπες μεθόδους είναι ο παθογόνος μικροοργανισμός και ότι δεν αποτελεί μέλος της φυσιολογικής χλωρίδας του στοματοφάρυγγα. Επίσης αποτελεί ίσως την γρηγορότερη και ευκολότερη μέθοδο για διαπίστωση του Streptococcus pneumonia ως αιτιολογικού παράγοντα της πνευμονίας. Συμπερασματικά διαφαίνεται ότι χρήση όλων των διαθέσιμων διαγνωστικών μεθόδων ανίχνευσης παθογόνων του κατώτερου αναπνευστικού, πλεονεκτεί έναντι της μεμονωμένης χρήσης μιας μόνο μεθόδου. Η χρήση όλων των μεθόδων οδηγεί σε πιο ασφαλή αποτελέσματα αφού μπορεί να διαπιστωθεί αν ο μικροοργανισμός που ανιχνεύθηκε αποτελεί τον παθογόνο μικροοργανισμό στον οποίο οφείλεται η πνευμονία και δεν αποτελεί απλώς μέλος της φυσιολογικής χλωρίδας. Επίσης με τη χρήση όλων των διαγνωστικών μεθόδων μπορεί να εξαχθεί ασφαλέστερο συμπέρασμα όσον αφορά στη συχνότητα των παθογόνων μικροοργανισμών που ενοχοποιούνται στην πρόκληση πνευμονίας της κοινότητας. Σε συνδυασμό όλες οι διαγνωστικές μέθοδοι αυξάνουν τον αριθμό των ασθενών με αιτιολογική διάγνωση της πνευμονίας και επομένως γίνεται πιο ασφαλής η αντιμετώπισή τους όσον αφορά στη σωστή επιλογή του αντιβιοτικού σχήματος.

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