Detection of Crimean-Congo hemorrhagic fever virus genome in saliva and urine.

BACKGROUND: The Crimean-Congo hemorrhagic fever (CCHF) virus is transmitted by tick bites and by contact with the blood or tissues of infected patients and livestock. This study was designed to investigate the genome of CCHF virus in saliva and urine samples of patients with CCHF. METHODS: Eight patients with laboratory-confirmed CCHF were included in the study. The diagnosis was made by detection of viral RNA in blood by real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR). Samples of saliva from six patients and samples of urine from three patients were collected at the same time as the blood samples and analyzed for viral RNA. RESULTS: The genome of CCHF virus was detected in the saliva from five of the six patients and in the urine from two of the three patients. The levels of viral load in the saliva and urine samples were similar to those in the blood samples in all but one patient, in whom higher levels were detected in blood compared to saliva or urine. CONCLUSIONS: This study shows that during human infection with CCHF virus, viral genomes are present in the saliva and urine. Further studies to isolate infectious viruses from these fluids and to study whether they represent an infectious risk are underway.

Presence of drug resistance in intestinal lactobacilli of dairy and human origin in Turkey.

The prevalence of different resistance genes was investigated in lactobacilli of human and dairy origin by PCR. The presence of erm, van, tet, and cat-TC genes were determined in 16 raw milk, 15 cream, 10 yogurt, 50 hand-made cheese, and 20 industrially produced white-cheese samples of dairy origin and 16 mouth, 32 fecal, and 36 vaginal samples from different subjects of human origin. Lactobacilli of dairy and human origin were found to carry only erm(B) and tet(M) genes. The majority of the isolates, Lactobacillus crispatus (61), Lactobacillus gasseri (49), Lactobacillus plantarum (80) studied were found to harbor either erm(B) or tet(M) gene or both. No resistant lactobacilli was found in raw-milk and cream samples. All the human fecal samples and the majority of vaginal (29 of 36) and mouth (10 of 14) samples were found to carry the resistance genes. While a third of the hand-made cheeses carried resistant lactobacilli only one industrially produced cheese was found to carry resistant lactobacilli. Furthermore, the genes were found in the non-starter species, Lactobacillus acidophilus and Lb. plantarum, indicating that industrially produced cheeses in this respect could be considered more favorable. These results indicate that drug resistance seems to be very common in Turkey. Even though the number of dairy samples harboring the resistance genes (17 of 111) is smaller in regards to human samples, 10% of them were still found to carry the resistance genes as well. The presence of the resistance genes in majority of the samples of human origin and in minority of the samples of dairy origin indicates that drug resistance may be acquired in the intestinal tract during passage and spread to dairy products by the hands of workers during production.

GJB2 gene mutations causing familial hereditary deafness in Turkey.

Mutations in Connexin 26 (Cx26) play an important role in autosomal non-syndromic hereditary hearing loss. In this study, our objective was to find out the significance of Cx26 mutations in Turkish families who had hereditary deafness. Fourteen families who had at least two prelingually deaf children per family were included in the study. One affected child from each of the 14 families was selected for single-stranded conformational polymorphism SSCP analysis. Three PCR reactions were used for each subject to amplify the entire Cx26 coding region with overlap. PCR products were sequenced on an Applied Biosystems (ABI) model 3700 automated sequencer. Six of the 14 representative family members (42.9%) demonstrated shifts on SSCP and were subsequently sequenced for Exons 1 and 2 of GJB2 and were tested for the 432 kb upstream deletion. No mutations were found in Exon 1 and no 432 kb deletions were noted. Three different GJB2 mutations were found in Exon 2 of the probands, which were 35delG, 299-300delAT, and 487G > A (M163V). GJB2 mutations were detected in 21.4% of the families. Two patients were homozygous for 35delG and 299-300delAT mutations, and were given a diagnosis of DFNB1 deafness (14.3%). Two different polymorphisms, 457G > A (V153I) and 380G > AG (R127H) were also found. In conclusion, although GJB2 mutations were detected in 21.4% of the families tested, only 14.3% of subject representatives were homozygous and therefore deafness caused by Cx26 mutation segregated with DFNB1. Thus, contribution of GJB2 mutations appears less significant in familial deafness. This necessitates further assessment for the other known gene regions as well as a search for new genetic factors in familial type of genetic deafness.

Evaluation of a polymerase chain reaction amplification method for Mycobacterium tuberculosis complex on samples from different sources.

Using primers specific for the IS6110 region of Mycobacterium tuberculosis complex, successful amplification by the polymerase chain reaction was demonstrated in 81 of 84 archive specimens from patients who had been clinically diagnosed 2 months to 16 y previously as having tuberculosis. Depending on the time of storage of the specimens, extra DNA bands were found in addition to the IS6110 region band.