Detection of Mycobacterium avium subsp. paratuberculosis in Cow Milk Using Culture and PCR methods

Mycobacterium avium subspecies paratuberculosis (MAP) is the cause of John’s disease also calledparatuberculosis. This is economically one of the important infectious diseases in cattle and ruminanthusbandry. This disease is manifested as granulomatosis entrocolitis, lymphadenitis and inflammation locallymphatic vessels. The typical sign of this disease is progressive loss of weight. Considering the importanceof detection of this disease in this study, two methods, culture and PCR, were used for the identification ofthis microorganism. In this study 100 milk samples from apparently healthy cows and 100 milk samplesfrom cows that have been suspicious of John’s disease were taken from in Sarab, East Azarbaijan, Iran.Direct microscope observation after ziehl-neelsen staining was done. Then, bacterial culture on specificmedium was carried out, and finally, identification of Mycobacterium avium subsp. paratuberculosis wasexamined using PCR and specific primers. Using direct observation, culture and PCR analyses showed thatfrom 100 healthy cow milk samples, 8, 9 and 12 samples were positive MAP for each method respectively.The results of direct observation, culture and PCR analysis on affected cows were 15, 40 and 44, respectively. The results of this study showed that culture and PCR analyses methods are important in the identification of the causes of this disease. Therefore, considering the frequency of the disease in the studied region, either of those methods can be used in the microorganism identification

Identification and determination of the prevalence of Toxoplasma gondii in patients with chronic renal failure by ELISA and PCR

Objective: To detect Toxoplasma gondii (T. gondii) among end-stage renal disease (ESRD) patients. Methods: This case-control study was conducted on 180 blood samples. In compliance with all ethical principles, 90 blood samples were taken from hemodialysis patients with ESRD and 90 samples from healthy volunteers. T. gondii screening was done using ELISA to search for immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies and by PCR for amplification of the T. gondii genome using specific primers. Results: The results were analyzed using SPSS software. Out of 90 patients on hemodialysis, 54 (60.0) were positive for anti-toxoplasma IgG antibody, 3 (3.3) for anti-toxoplasma IgM antibody and 5 patients (6) were positive by PCR. From 90 healthy volunteers, 34 (37.8) were positive for anti-toxoplasma IgG antibody. All the healthy volunteers were negative for anti-toxoplasma IgM antibody and in PCR. Compared with the gold standard method of ELISA, PCR had 100 sensitivity and 98.9 specificity in detection of T. gondii. Conclusions: PCR alongside serologic methods can be valuable for T. gondii screening. Given the high prevalence of T. gondii among hemodialysis patients with ESRD, T. gondii screening together with sanitary control of biological agents was recommended in dialysis units. © 2016 Asian Pacific Tropical Medicine Press

Lethal Pneumocystis jiroveci pneumonia 24 Years After Kidney Transplantation

INTRODUCTION: Pneumocystis jiroveci is an opportunistic infectious fungus in immunosuppressed patients, particularly in ones with acquired immunodeficiency syndrome (AIDS). The use of immunosuppressive drugs especially corticosteroids predisposes the transplanted patients to a variety of infectious diseases including Pneumocystis infection. In many developed countries, the incidence of Pneumocystis jiroveci pneumonia (PJP) is dwindling in transplant patients receiving appropriate prophylaxis. In this study, definitive diagnosis of Pneumocystis infection in a patient receiving kidney transplant was presented. CASE PRESENTATION: The patient was a 45-year-old man with a history of kidney transplantation 24 years ago, admitted to a specialized hospital in Tehran because of fever and respiratory distress. Upon admission, the patient showed symptoms of unconsciousness and shortness of breath. Paraclinical tests and complementary examinations such as microscopic observation and molecular analysis confirmed the definitive diagnosis of Pneumocystis infection. Specific treatment with trimethoprim/sulfamethoxazole was carried out alongside other therapeutic measures; but unfortunately the patient did not respond to the specific treatment and died in the course of a progressive disease. DISCUSSION: The disease progress in these patients can still be fast and deadly. Applying rapid molecular diagnostic techniques to start appropriate and timely treatment is essential. Utilization of such diagnostic methods is recommended in our country