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Sperm Banking
Published in Botros Rizk, Ashok Agarwal, Edmund S. Sabanegh, Male Infertility in Reproductive Medicine, 2019
Rakesh Sharma, Alyssa M. Giroski, Ashok Agarwal
Men who are seropositive for human immunodeficiency virus (HIV) and are coinfected with hepatitis C virus (HCV) have demonstrated the presence of HIV and HCV in the ejaculate and risk sexual transmission [70]. Nested polymerase chain reaction (nPCR) is the most sensitive method of detecting HIV and HCV [71,72]. Sperm-washing procedures can eliminate the viruses from the ejaculate and their absence can be confirmed by nPCR. Only those samples that are confirmed negative for HIV and HCV are subsequently used in ARTs with a negligible risk of transmission.
Histopathology
Published in Peter D O Davies, Stephen B Gordon, Geraint Davies, Clinical Tuberculosis, 2014
Two immunohistochemical antibodies, M. tuberculosis complex specific antibody (anti-MPT64) and BCG antibody, have been tested on pleural biopsies for diagnosis and species identification [18]. They were compared with a ZN stain and nested polymerase chain reaction (N-PCR). Sensitivity was 81% and 56%, respectively; specificity was 100% and 78%, respectively. PCR sensitivity was 80% and specificity was 100%. These techniques are expensive and not suitable for resource-poor settings.
Cystoisospora belli
Published in Dongyou Liu, Laboratory Models for Foodborne Infections, 2017
Chaturong Putaporntip, Somchai Jongwutiwes
Amplification of a specific region in the small subunit ribosomal RNA gene (18S rDNA) of C. belli by nested polymerase chain reaction (PCR) has been successfully applied to duodenal biopsy and bile and stool samples without cross-reactivity to other enteric protozoa [8,61]. PCR targeting the 5.8S ribosomal RNA gene and the internal transcribed spacer II (ITS2) region is proven to be more sensitive than microscopy in diagnosing C. belli [62]. Likewise, quantitative PCR assay with melting curve analysis using primers derived from the 18S rDNA can differentiate human enteric coccidian protozoa [63]. Diagnosis of C. belli infection by PCR involves the isolation of C. belli DNA from clinical specimens that can be stool, duodenal fluid, tissue sample, or other potential sources of infections. The source of C. belli DNA is usually from oocyst stage in stool, while tissue sample contains any stages of endogenous development. Freshly collected stool specimen, frozen stool sample, or stool preserved in 80% ethanol can be used for extraction of C. belli DNA. Ethanol in preserved stool samples should be removed before disruption of oocysts. Duodenal fluid or tissue samples can be kept at −20°C, and ethanol should not be used for preservation of these samples. Because the oocyst wall of coccidian protozoa is solid and resistant to diverse environmental conditions, it is recommended that the oocyst wall should be ruptured so that internal contents, either sporoblasts or sporozoites, are exposed to DNA extraction solution. Examples of simple procedures to disrupt oocyst wall of C. belli are mechanical disruption and freeze-thaw method.
Presumed Tuberculous Sclerokeratitis Presenting with Hypopyon
Published in Ocular Immunology and Inflammation, 2019
On examination, best corrected visual acuity (BCVA) in her right and left eye was 6/12 and 6/6 respectively. Slit-lamp examination of the right eye showed marked hyperemia, multiple scleral nodules superiorly with diffuse congestion of the deeper episcleral vessels and anterior chamber reaction- cells 2+, Flare 1+ with hypopyon. Two crescent-shaped white-grayish stromal opacities were seen 2 mm from the limbus in inferior cornea. (Figure 1a) Slit-lamp and fundus examination of the left eye was unremarkable. Intraocular pressure measured with Goldman applanation tonometry in her right and left eye was 14 and 12 mm of Hg. She was started on frequent topical steroid (Prednisolone acetate suspension, 1% one hourly) and atropine eye drop three times a day. Her laboratory investigation revealed negative serology for syphilis, normal angiotensin-converting enzyme, negative human leucocyte antigen (HLA) B27, antinuclear antibody, rheumatoid factor, cytoplasmic, and perinuclear antineutrophil cytoplasmic antibodies. Nested polymerase chain reaction (PCR) from aqueous aspirate from anterior chamber paracentesis of her right eye was found to be negative for MPB 64 and IS 6110 genes. Her interferon gamma release assay (QuantiFERON®-TB) was positive and high-resolution computerized tomography (HRCT) of chest revealed right hilar lymphadenopathy. After consultation from a chest physician and in-house physician, she was started on anti-tubercular therapy (ATT) and oral steroid (1 mg/kg/day in tapering doses).
Culture and Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) Proven Mycobacterium Tuberculosis Endophthalmitis: A Case Series
Published in Ocular Immunology and Inflammation, 2018
Ekta Rishi, Pukhraj Rishi, K. Lily Therese, Gayathri Ramasubban, Jyotirmay Biswas, Tarun Sharma, Pramod Bhende, Pradeep Susvar, Mamta Agarwal, Amala Elizabeth George, Kushal Delhiwala, Vishal Rajan Sharma
Nested polymerase chain reaction (nPCR) using primers targeting MPB 64 and IS6110 genes of MTB are performed as described previously.9 Real-time polymerase chain reaction (rt-PCR) for MTB is carried out using Genosen’s MTB complex quantitative rt-PCR kit (Genome Diagnostics, Parwanoo, India). Rt-PCR for quantification of MTB DNA is carried out as a 25-μl reaction, using 12 μl of MTB complex supermix R1, 2.5 μl of magnesium solution R2, 0.5 μl of internal control IC 1 R3,and 10 μl of MTB DNA. The amplification was carried out with initial denaturation at 95ºC for 10 min, followed by 45 cycles of 95ºC for 15 s, 60ºC for 20 s, and 72ºC for 15 s. Quantitation analysis for the internal control and MTB DNA is carried out using JOE (yellow) and FAM (green) channels. The copy number of M. tuberculosis is expressed in copies per ml of MTB DNA.
Molecular surveillance of chloroquine drug resistance markers (Pfcrt and Pfmdr1) among imported Plasmodium falciparum malaria in Qatar
Published in Pathogens and Global Health, 2018
Anushree Acharya, Devendra Bansal, Praveen K. Bharti, Fahmi Y. Khan, Salem Abusalah, Ashraf Elmalik, Mohammed ElKhalifa, Pradyumna K. Mohapatra, Jagadish Mahanta, Rakesh Sehgal, Neeru Singh, Ali A. Sultan
Intravenous blood samples, in sterile vacutainer, were collected from the patients attending the Hamad General Hospital, HMC, Qatar. The inclusion criteria included signs and symptoms of acute uncomplicated malaria (according to WHO), patients with P. falciparum asexual parasite without severe anemia [1]. Pregnant women, patients with signs and symptoms of severe and complicated malaria (as defined by WHO) [1], and patients with severe anemia or if they have second major illness, were excluded from this study. All the samples were first screened for malaria parasites by microscopy using Giemsa staining and confirmed by nested polymerase chain reaction (PCR). All subjects were interviewed using a structured questionnaire to collect socio-demographics and risk factors such as travel history, blood transfusions. Other data from medical records include age, nationality, clinical history, parasitaemia and hemoglobin were collected from each subject. Patients were given treatment as per the Ministry of Public Health Qatar.