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Prevalence and Distribution of BK virus Subtypes in Renal Transplant Recipients Referred to Golestan Hospital in Ahvaz, Iran.

Kaydani GA, Makvandi M, Samarbafzadeh A, Shahbazian H, Hamidi Fard M - Jundishapur J Microbiol (2015)

Bottom Line: Forty-eight cases (94.11%) were subtype I and 3 others (5.89%) were subtype IV using the RFLP method.None of the patient's urine samples were positive for subtypes II and III.This helps better transplantation result and may prevent graft rejection.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, Infectious and Tropical Diseases Research Center, Faculty of Paramedical, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.

ABSTRACT

Background: BK virus (BKV) belongs to the human Polyomaviridae and the primary BKV infection is occurred during childhood then the virus could be latent through life, especially in the kidneys and urinary system. It became reactive after an immunocompromised status, such as pregnancy or transplantation. Isolated BKV from different locations of the world is grouped into four subtypes using serological and genotyping methods. The BKV subtype I is the dominant one and has worldwide distribution.

Objectives: According to our knowledge, there are no data about the BKV prevalence and its genotypes in southwest part of Iran. Considering the high prevalence of renal failure and kidney transplant patients in this part, and the role of BKV in graft rejection, this study aimed to determine the prevalence of BKV infection in renal transplant recipients referred to Golestan Hospital in Ahvaz City, Iran.

Patients and methods: Urine samples were collected from 122 kidney transplant recipients referred to Golestan Hospital in Ahvaz, southwest of Iran. The extracted DNA was amplified by Polymerase Chain Reaction, and subtype of each positive sample was determined using Restriction Fragment Length Polymorphism (RFLP) and sequencing methods.

Results: From all study population, 51/122 (41.8%) urine samples were positive for BKV DNA and the other samples were negative (71/122). Forty-eight cases (94.11%) were subtype I and 3 others (5.89%) were subtype IV using the RFLP method. None of the patient's urine samples were positive for subtypes II and III.

Conclusions: Our work is the second study in Iran and considering huge numbers of transplantation in Iran and Khuzestan Province, south western of Iran, in addition to the role of this virus in kidney transplant rejection, routine evaluation of BKV positivity is recommended both for graft recipient and donors. This helps better transplantation result and may prevent graft rejection.

No MeSH data available.


Related in: MedlinePlus

Polymerase Chain Reaction Products for Positive and Negative Samples on 1.7% Agarose GelP, positive control; N, negative control; L, 100 bp; 1-4, positive samples; 5-6, negative samples
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fig18075: Polymerase Chain Reaction Products for Positive and Negative Samples on 1.7% Agarose GelP, positive control; N, negative control; L, 100 bp; 1-4, positive samples; 5-6, negative samples

Mentions: Extracted DNA was amplified using 5 μL DNA template, 0.3 μL Taq Polymerase, 1 μL dNTP, 0.25 μL each forward and reveres primers, 2 μL MgCl2 and 5 μL 10X PCR buffer, all mixed in a microtube and the ultimate volume reach to 50 μL using nuclease-free distilled water. For each run, we used Diethylpyrocarbonate (DEPC) water as a negative control and confirmed positive sample DEPC water as a negative control and confirmed positive sample from our previous study (9) as a positive control. After activation of reaction in 95°C for 5 minutes, amplification was performed for 50 cycles. Cycle program was as: 94°C for 60, 55°C for 60 and 72°C for 90 seconds. A final extension 72°C for 5 minutes was the ultimate step of cycles. Polymerase chain reactions (PCRs) were performed using the Peqlab thermocycler (Peq star, 96 Universal Gradient -Germany). The 342 bp amplification product was detected using 1.7% gel agarose socked in ethidium bromide (Figure 1). Positive samples for BKV were sent to Noor Genetic Laboratory in Ahvaz for sequencing. Sequences were analyzed using the neighbor Toning phylogenetic tree software and compared by reference ones.


Prevalence and Distribution of BK virus Subtypes in Renal Transplant Recipients Referred to Golestan Hospital in Ahvaz, Iran.

Kaydani GA, Makvandi M, Samarbafzadeh A, Shahbazian H, Hamidi Fard M - Jundishapur J Microbiol (2015)

Polymerase Chain Reaction Products for Positive and Negative Samples on 1.7% Agarose GelP, positive control; N, negative control; L, 100 bp; 1-4, positive samples; 5-6, negative samples
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4385253&req=5

fig18075: Polymerase Chain Reaction Products for Positive and Negative Samples on 1.7% Agarose GelP, positive control; N, negative control; L, 100 bp; 1-4, positive samples; 5-6, negative samples
Mentions: Extracted DNA was amplified using 5 μL DNA template, 0.3 μL Taq Polymerase, 1 μL dNTP, 0.25 μL each forward and reveres primers, 2 μL MgCl2 and 5 μL 10X PCR buffer, all mixed in a microtube and the ultimate volume reach to 50 μL using nuclease-free distilled water. For each run, we used Diethylpyrocarbonate (DEPC) water as a negative control and confirmed positive sample DEPC water as a negative control and confirmed positive sample from our previous study (9) as a positive control. After activation of reaction in 95°C for 5 minutes, amplification was performed for 50 cycles. Cycle program was as: 94°C for 60, 55°C for 60 and 72°C for 90 seconds. A final extension 72°C for 5 minutes was the ultimate step of cycles. Polymerase chain reactions (PCRs) were performed using the Peqlab thermocycler (Peq star, 96 Universal Gradient -Germany). The 342 bp amplification product was detected using 1.7% gel agarose socked in ethidium bromide (Figure 1). Positive samples for BKV were sent to Noor Genetic Laboratory in Ahvaz for sequencing. Sequences were analyzed using the neighbor Toning phylogenetic tree software and compared by reference ones.

Bottom Line: Forty-eight cases (94.11%) were subtype I and 3 others (5.89%) were subtype IV using the RFLP method.None of the patient's urine samples were positive for subtypes II and III.This helps better transplantation result and may prevent graft rejection.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, Infectious and Tropical Diseases Research Center, Faculty of Paramedical, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.

ABSTRACT

Background: BK virus (BKV) belongs to the human Polyomaviridae and the primary BKV infection is occurred during childhood then the virus could be latent through life, especially in the kidneys and urinary system. It became reactive after an immunocompromised status, such as pregnancy or transplantation. Isolated BKV from different locations of the world is grouped into four subtypes using serological and genotyping methods. The BKV subtype I is the dominant one and has worldwide distribution.

Objectives: According to our knowledge, there are no data about the BKV prevalence and its genotypes in southwest part of Iran. Considering the high prevalence of renal failure and kidney transplant patients in this part, and the role of BKV in graft rejection, this study aimed to determine the prevalence of BKV infection in renal transplant recipients referred to Golestan Hospital in Ahvaz City, Iran.

Patients and methods: Urine samples were collected from 122 kidney transplant recipients referred to Golestan Hospital in Ahvaz, southwest of Iran. The extracted DNA was amplified by Polymerase Chain Reaction, and subtype of each positive sample was determined using Restriction Fragment Length Polymorphism (RFLP) and sequencing methods.

Results: From all study population, 51/122 (41.8%) urine samples were positive for BKV DNA and the other samples were negative (71/122). Forty-eight cases (94.11%) were subtype I and 3 others (5.89%) were subtype IV using the RFLP method. None of the patient's urine samples were positive for subtypes II and III.

Conclusions: Our work is the second study in Iran and considering huge numbers of transplantation in Iran and Khuzestan Province, south western of Iran, in addition to the role of this virus in kidney transplant rejection, routine evaluation of BKV positivity is recommended both for graft recipient and donors. This helps better transplantation result and may prevent graft rejection.

No MeSH data available.


Related in: MedlinePlus