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Impact of RB1 gene screening from blood collected on a single day from 411 family members of 113 Retinoblastoma survivors in India



To analyse the profile and implication of genetic testing in a cohort of retinoblastoma (RB) patients and their families conducted on a single day during World Retinoblastoma Awareness Week 2017.


Retrospective analysis of blood samples were collected from 411 subjects, including 113 probands at a camp organised for RB awareness and were analysed for RB1 mutations by Sanger sequencing and Multiplex Ligation-dependent Probe Amplification (MLPA). If germline mutations were detected, the parents and siblings of the proband were tested for the same mutation.


Germline RB1 mutations were identified in 61/113(54%) probands with a mutation detection rate of 96% (47/49) and 22% (14/64) for bilateral and unilateral RB, respectively. Ten novel pathogenic mutations were identified. Splice mutation was most common (31%) followed by nonsense mutation (26%). The mean age at RB diagnosis was significantly lower in patients having germline RB1 mutation (mean 10.7 months ±2.5) compared to those without (mean 27.2 months ±6.5) (p = <0.0001). Parental transmission of the mutant allele was detected in 15/61(25%) cases of which 11(18%) parents were unaffected indicating incomplete penetrance. The origin of the variant allele was both paternal (n = 7) and maternal (n = 4) wherein 5 were bilateral and 6 unilateral.


The detection of a germline mutation impacts the proband and family members due to its implications on change in prognosis, frequency of subsequent evaluations, screening for ocular and non-ocular cancers, and surveillance of family and future progeny.

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Fig. 1: Distribution and schematic representation of mutations across RB1 gene.
Fig. 2: Representative pedigree of families with low penetrance RB1 mutation.
Fig. 3: Post testing surveillance in familial RB.

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Data availability

The data supporting this study’s findings are available from the corresponding author upon request.


  1. Knudson AG. Mutation and cancer: statistical study of retinoblastoma. Proc Natl Acad Sci USA. 1971;68:820–3.

    Article  ADS  PubMed  PubMed Central  Google Scholar 

  2. Soliman SE, Racher H, Zhang C, MacDonald H, Gallie BL. Genetics and molecular diagnostics in Retinoblastoma-An Update. Asia Pac J Ophthalmol. 2017;6:197–207.

    CAS  Google Scholar 

  3. Athavale V, Khetan V. Knudson to embryo selection: A story of the genetics of retinoblastoma. Taiwan J Ophthalmol. 2018;8:196–204.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Dryja TP, Mukai S, Petersen R, Rapaport JM, Walton D, Yandell DW. Parental origin of mutations of the retinoblastoma gene. Nature. 1989;339:556–8.

    Article  ADS  CAS  PubMed  Google Scholar 

  5. Zhu XP, Dunn JM, Phillips RA, Goddard AD, Paton KE, Becker A, et al. Preferential germline mutation of the paternal allele in retinoblastoma. Nature. 1989;340:312–3.

    Article  ADS  CAS  PubMed  Google Scholar 

  6. Chawla B, Hasan F, Azad R, Seth R, Upadhyay AD, Pathy S, et al. Clinical presentation and survival of retinoblastoma in Indian children. Br J Ophthalmol. 2016;100:172–8.

    Article  PubMed  Google Scholar 

  7. Shah PK, Narendran V, Kalpana N. Outcomes of intra- and extraocular retinoblastomas from a single institute in South India. Ophthalmic Genet. 2015;36:248–50.

    Article  PubMed  Google Scholar 

  8. Francis JH, Roosipu N, Levin AM, Brodie SE, Dunkel IJ, Gobin YP, et al. Current treatment of bilateral retinoblastoma: the impact of intraarterial and intravitreous chemotherapy. Neoplasia. 2018;20:757–63.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Linn Murphree A. Intraocular retinoblastoma: the case for a new group classification. Ophthalmol Clin North Am. 2005;18:41–53. viii

    Article  CAS  PubMed  Google Scholar 

  10. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988;16:1215.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Thirumalairaj K, Abraham A, Devarajan B, Gaikwad N, Kim U, Muthukkaruppan V, et al. A stepwise strategy for rapid and cost-effective RB1 screening in Indian retinoblastoma patients. J Hum Genet. 2015;60:547–52.

    Article  PubMed  Google Scholar 

  12. Tomar S, Sethi R, Sundar G, Quah TC, Quah BL, Lai PS. Mutation spectrum of RB1 mutations in retinoblastoma cases from Singapore with implications for genetic management and counselling. PLoS One. 2017;12:e0178776.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Shahraki K, Ahani A, Sharma P, Faranoush M, Bahoush G, Torktaz I, et al. Genetic screening in Iranian patients with retinoblastoma. Eye. 2017;31:620–7.

    Article  CAS  PubMed  Google Scholar 

  14. Lan X, Xu W, Tang X, Ye H, Song X, Lin L, et al. Spectrum of RB1 germline mutations and clinical features in Unrelated Chinese patients With Retinoblastoma. Front Genet. 2020;11:142.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Sagi M, Frenkel A, Eilat A, Weinberg N, Frenkel S, Pe’er J, et al. Genetic screening in patients with Retinoblastoma in Israel. Fam Cancer. 2015;14:471–80.

    Article  CAS  PubMed  Google Scholar 

  16. Mohd Khalid MKN, Yakob Y, Md Yasin R, Wee Teik K, Siew CG, Rahmat J, et al. Spectrum of germ-line RB1 gene mutations in Malaysian patients with retinoblastoma. Mol Vis. 2015;21:1185–90.

    PubMed  PubMed Central  Google Scholar 

  17. Rushlow D, Piovesan B, Zhang K, Prigoda-Lee NL, Marchong MN, Clark RD, et al. Detection of mosaic RB1 mutations in families with retinoblastoma. Hum Mutat. 2009;30:842–51.

    Article  CAS  PubMed  Google Scholar 

  18. Lohmann DR, Gallie BL. Retinoblastoma: revisiting the model prototype of inherited cancer. Am J Med Genet C Semin Med Genet. 2004;129C:23–28.

    Article  PubMed  Google Scholar 

  19. Harbour JW. Molecular basis of low-penetrance retinoblastoma. Arch Ophthalmol. 2001;119:1699–704.

    Article  CAS  PubMed  Google Scholar 

  20. Lohmann DR, Brandt B, Höpping W, Passarge E, Horsthemke B. Distinct RB1 gene mutations with low penetrance in hereditary retinoblastoma. Hum Genet. 1994;94:349–54.

    Article  CAS  PubMed  Google Scholar 

  21. Ketteler P, Hülsenbeck I, Frank M, Schmidt B, Jöckel K-H, Lohmann DR. The impact of RB1 genotype on incidence of second tumours in heritable retinoblastoma. Eur J Cancer. 2020;133:47–55.

    Article  CAS  PubMed  Google Scholar 

  22. Shah PK, Sripriya S, Narendran V, Pandian AJ. Prenatal genetic diagnosis of retinoblastoma and report of RB1 gene mutation from India. Ophthalmic Genet. 2016;37:430–3.

    Article  CAS  PubMed  Google Scholar 

  23. Soliman SE, Dimaras H, Khetan V, Gardiner JA, Chan HSL, Héon E, et al. Prenatal versus postnatal screening for familial retinoblastoma. Ophthalmology. 2016;123:2610–7.

    Article  PubMed  Google Scholar 

  24. Akolekar R, Beta J, Picciarelli G, Ogilvie C, D’Antonio F. Procedure-related risk of miscarriage following amniocentesis and chorionic villus sampling: a systematic review and meta-analysis. Ultrasound Obstet Gynecol. 2015;45:16–26.

    Article  CAS  PubMed  Google Scholar 

  25. Xu K, Rosenwaks Z, Beaverson K, Cholst I, Veeck L, Abramson DH. Preimplantation genetic diagnosis for retinoblastoma: the first reported liveborn. Am J Ophthalmol. 2004;137:18–23.

    Article  PubMed  Google Scholar 

  26. Gerrish A, Bowns B, Mashayamombe-Wolfgarten C, Young E, Court S, Bott J, et al. Non-invasive prenatal diagnosis of retinoblastoma inheritance by combined targeted sequencing strategies. J Clin Med. 2020;9:3517.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Manjandavida FP, Xia J, Zhang J, Tang XY, Yi HR. In-utero ultrasonography detection of fetal retinoblastoma and neonatal selective ophthalmic artery chemotherapy. Indian J Ophthalmol. 2019;67:958–60.

    Article  PubMed  PubMed Central  Google Scholar 

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We thank all our patients who participated in our study. We would also like to thank Sun Foundation and our colleagues in the department of vitreoretinal and molecular genetics for providing their support.

Presentations Part of this study was presented at the International Society of Genetic Eye Diseases and Retinoblastoma, Giessen, Germany, August 29–31, 2019.


No funding was received for this study.

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Authors and Affiliations



Conceptualisation- PKS and AV, data curation-PKS, PM and AV, data analysis- PM, KKS and AV, paper writing- PM, KKS, PKS and AV. All authors accepted the final draft of the paper.

Corresponding author

Correspondence to Parag K. Shah.

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The authors declare no competing interests.

Ethics statement

This study was approved by the Institutional Human Ethics Committee, PSG IMS&R (Project number 17/109).

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Vanniarajan, A., Maitra, P., Saraswathi, K.K. et al. Impact of RB1 gene screening from blood collected on a single day from 411 family members of 113 Retinoblastoma survivors in India. Eye (2024).

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