Search results for: Mehrdad Jalali

Authors: Mehrdad Hashemi, Mitra Behrooz Aghdam, Reza Mahdian, Ahmad Reza Kamyab

Abstract:

Cytogenetic analysis still remains the gold standard method for prenatal diagnosis of trisomy 21 (Down syndrome, DS). Nevertheless, the conventional cytogenetic analysis needs live cultured cells and is too time-consuming for clinical application. In contrast, molecular methods such as FISH, QF-PCR, MLPA and quantitative Real-time PCR are rapid assays with results available in 24h. In the present study, we have successfully used a novel MGB TaqMan probe-based real time PCR assay for rapid diagnosis of trisomy 21 status in Down syndrome samples. We have also compared the results of this molecular method with corresponding results obtained by the cytogenetic analysis. Blood samples obtained from DS patients (n=25) and normal controls (n=20) were tested by quantitative Real-time PCR in parallel to standard G-banding analysis. Genomic DNA was extracted from peripheral blood lymphocytes. A high precision TaqMan probe quantitative Real-time PCR assay was developed to determine the gene dosage of DSCAM (target gene on 21q22.2) relative to PMP22 (reference gene on 17p11.2). The DSCAM/PMP22 ratio was calculated according to the formula; ratio=2 -ΔΔCT. The quantitative Real-time PCR was able to distinguish between trisomy 21 samples and normal controls with the gene ratios of 1.49±0.13 and 1.03±0.04 respectively (p value <0.001). These results represent the presence of 3 copies of target gene in DS samples Vs 2 copies in normal controls. The results of quantitative Real-time PCR were in complete agreement with results of cytogenetic analysis. This study confirms previous reports regarding successful implementation of quantitative Real-time PCR for detection of trisomy 21. However, the assay has been improved by using MGB probes and more accurate data analysis. This assay, in particular, when performed in combination with another molecular assay such as QF-PCR or MLPA, can be used as a reliable technique for rapid prenatal diagnosis of trisomy 21.

Keywords: Trisomy 21, Real-time PCR, MGB-TaqMan Probes, Gene Dosage.

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Authors: Mehrdad Ahmadi, Mohamad Babaie, Shiva Osouli, Bahareh Salehi, Nadia Kalantaraian

Abstract:

The olive fruit fly, Bactrocera oleae Gmelin (Diptera: Tephritidae), is one of the most serious pests in olive orchards in growing province in Iran. The female lay eggs in green olive fruit and larvae hatch inside the fruit, where they feed upon the fruit matters. One of the main ecologically friendly and species-specific systems of pest control is the sterile insect technique (SIT) which is based on the release of large numbers of sterilized insects. The objective of our work was to develop a SIT against B. oleae by using of gamma radiation for the laboratory and field trial in Iran. Oviposition of female mated by irradiated males is one of the main parameters to determine achievement of SIT. To conclude the sterile dose, pupae were placed under 0 to 160 Gy of gamma radiation. The main factor in SIT is the productivity of females which are mated by irradiated males. The emerged adults from irradiated pupae were mated with untreated adults of the same age by confining them inside the transparent cages. The fecundity of the irradiated males mated with non-irradiated females was decreased with the increasing radiation dose level. It was observed that the number of eggs and also the percentage of the egg hatching was significantly (P < 0.05) affected in either IM x NF crosses compared with NM x NF crosses in F₁ generation at all doses. Also, the statistical analysis showed a significant difference (P < 0.05) in the mean number of eggs laid between irradiated and non-irradiated females crossed with irradiated males, which suggests that the males were susceptible to gamma radiation. The egg hatching percentage declined markedly with the increase of the radiation dose of the treated males in mating trials which demonstrated that egg hatch rate was dose dependent. Our results specified that gamma radiation affects the longevity of irradiated B. oleae larvae (established from irradiated pupae) and significantly increased their larval duration. Results show the gamma radiation, and SIT can be used successfully against olive fruit flies.

Keywords: Fertility, olive fruit fly, radiation, SIT.

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Authors: Shiva Osouli, Maryam Atapour, Mehrdad Ahmadi, Shima Shokri

Abstract:

Citrus leaf miner (Phyllocnistis citrella Stainton) is native to Asia and one of the most serious pests of Iran’s citrus nursery stocks. In the present study, the possibility of insect mass rearing on four various citrus hosts and the effects of gamma irradiation on the pupal mortality and reproduction of this pest were studied. Trifoliate orange and grapefruit showed less infection, while the number of pupae in Valencia oranges and sweet lemons cages was so high. There was not any significant difference between weight of male and female pupae among different citrus hosts, but generally the weight of male pupae was less than females. Use of Valencia orange or sweet lemons seedlings in especial dark emergence and oviposition cages could be recommended for mass rearing of this pest. In this study, the effects of gamma radiation at doses 100 to 450 Gy on biological and reproductive parameters of the pest has been determined. The results show that mean percent of pupal mortality increased with increasing doses and reached to 28.67% at 450 Gy for male pupae and 38.367% for female pupae. Also, the mean values of this parameter were higher for irradiated female, which indicated the higher sensitivity of this sex. The gamma ray irradiation from 200 and 300 Gy caused decrease in male and female adult moth longevity, respectively. The eggs were laid by emerged females, and their hatchability was decreased by increasing gamma doses. The fecundity of females in both combinations of crosses (irradiated male × normal female and irradiated female × normal male) did not differ, but fertility of laid eggs by irradiated female × normal male affected seriously and the mean values of this parameter reached to zero at 300 Gy. The hatchability percentage of produced eggs by normal female × irradiated male at 300 Gy was 23.29% and reached to less than 2 % at 450 Gy as the highest tested dose. The results of this test show that females have more radio-sensitivity in comparison to males.

Keywords: Citrus leaf miner, Phyllocnistis citrella, citrus hosts, mass rearing, sterile insect technique.

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