Salih Kuk

Stool sample storage conditions for the preservation of Giardia intestinalis DNA

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

Mikrosporidialar ve Mikrosporidiyozis

All microsporidia are obligate parasites and have no active stages outside their host cells. Microsporidia lack some typical eukaryotic characteristics. There are now over 1200 species identified in 144 genera. The most familiar stage of microsporidia is the small, highly resistant spore, the size of which differs according to the species and is often 1-10 μm. The general life cycle pattern of the microsporidia can be divided into three phases: the infective or environmental phase, the proliferative phase, and the sporogony or spore-forming phase. There are several methods for diagnosing microsporidia: light microscopic, transmission electron microscopy (TEM), immunofluorescence assays (IFA) and molecular methods. The clinical course of microsporidiosis depends on the immune status of the host and site of infection. Microsporidia can cause infections such as diarrhoea, keratitis, myositis, bronchitis and brochiolitis. Human microsporidiosis represents an important and rapidly emerging opportunistic disease, occurring mainly, but not exclusively, in severely immunocompromised patients with AIDS. The treatment of microsporidiosis is generally achieved with medications and supportive care. Depending on the site of infection and the microsporidia species involved, different medications are utilized. The most commonly used medications for microsporidiosis include albendazole and fumagillin.

[Investigation of anti-Echinococcus granulosus antibodies in patients with suspected cystic echinococcosis].

OBJECTIVE Cystic echninococcosis (CE) is an important helmintho-zoonotic disease causing health-threatening and economic losses for developing countries. In this study, anti-Echinococcus granulosus antibodies were evaluated in 1556 CE suspected patients (701 males, 855 females) who applied to the serology laboratory of the Parasitology Department of Erciyes University between June 1999 and July 2010. METHODS Fifty-six (3.6%) patients were evaluated with the three different methods of Indirect Hemagglutination Test (IHA), Indirect Fluorescent Antibody Test (IFAT) and Western blot (WB). 378 (24.3%) were tested with both IHA and IFAT, 123 (7.9%) with both IHA and WB,and 999 (64.2%) were evaluated with one of these three methods. RESULTS In 353 (22.7%) patients, anti-E. granulosus antibodies detected by one of above three methods were considered as positive. CONCLUSION Since some patients were assessed either as negative or positive with one of above test, we believe that it should be safer to use at least two tests together for diagnosis of CE.

The Epidemiology of Malaria in Kayseri Between 2001 and 2013

OBJECTIVE Malaria is the primary parasitic cause of morbidity and mortality in the world. As a result of the expansion of international travel in recent years, imported malaria cases especially are increasing in our country. Likewise, while there were more domestic cases earlier in Kayseri, more imported cases were seen in recent years. In our study, the epidemiology of malaria cases between the years of 2001-2013 is intended to be done with the data obtained from the Provincial Health Directorate. METHODS The data was performed retrospectively. RESULTS Considering the last 12 years of data; a total of 34,459 blood samples were analyzed and 47 of these cases were found to be malaria, 21 cases were domestic and others were imported cases of malaria. P. vivax was detected in all domestic cases. While one of the imported cases have been identified as P. malariae, others were P. falciparum. CONCLUSION We believe that our study of the epidemiological data would be beneficial for taking preventive cautions and fight against malaria.

Cryptosporidium parvum Gastroenteritis in a Patient with Renal Transplantation

In this study, a case who starting abundant watery diarrhea on the 14th day of renal transplantation is presented. Stool sample was analyzed for Cryptosporidium spp. by carbol fuchsin staining method, copro-ELISA and nested polimeraze chain reaction (PCR). From sample found positive by Carbol-fuchsin staining method and Copro-ELISA, DNA sequence analysis was performed, gel-purified from amplicon obtained by nested PCR. As a result of DNA sequence analysis was determined to be Cryptosporidium parvum. Although C. parvum is a rare causative agent of gastroenteritis it can be cause serious clinical diarrhea solid organ transplantation patient. As a result, also C.parvum must be considered as a causative agent of diarrhea occurring after organ transplantation.